NZ520798A - Tocopherols, tocotrienols, other chroman and side chain derivatives and uses thereof - Google Patents

Tocopherols, tocotrienols, other chroman and side chain derivatives and uses thereof

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Publication number
NZ520798A
NZ520798A NZ520798A NZ52079801A NZ520798A NZ 520798 A NZ520798 A NZ 520798A NZ 520798 A NZ520798 A NZ 520798A NZ 52079801 A NZ52079801 A NZ 52079801A NZ 520798 A NZ520798 A NZ 520798A
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New Zealand
Prior art keywords
4alkylene
compound
mmol
ether
cancer
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NZ520798A
Inventor
Shenquan Liu
Karen Israel
Robert G Sanders
Kimberly Kline
Laurence Hurley
Robb Gardner
Marla Menchaca
Weiping U
Puthucode N Ramanan
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Res Dev Foundation
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Priority claimed from US09/502,592 external-priority patent/US6770672B1/en
Application filed by Res Dev Foundation filed Critical Res Dev Foundation
Publication of NZ520798A publication Critical patent/NZ520798A/en

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    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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Abstract

An antiproliferative compound of structural formula (I) is disclosed, wherein X is oxygen, nitrogen or sulfur; Y is oxygen or NR6 , wherein the variables R1 to R6 are as defined in the specification. These compounds are useful for inducing apoptosis in a cell for the treatment of cell proliferative disease is selected from the group consisting of neoplastic diseases and non-neoplastic disorders.

Description

<div class="application article clearfix" id="description"> <p class="printTableText" lang="en">520798 <br><br> WO 01/58889 PCT/US01/04168 <br><br> TOCOPHEROLS, TOCOTRIENOLS, OTHER CHROMAN AND SIDE CHAIN DERIVATIVES AND USES THEREOF <br><br> 5 <br><br> BACKGROUND OF THE INVENTION <br><br> Fip.lri of the Invention 10 The present invention relates generally to the fields of organic chemistry and antiproliferative and pro-apoptotic compounds. More specifically, the present invention relates to chroman-based compounds and derivatives and analogs thereof, and their uses as cell anti-proliferative, proapoptotic, 15 immunomodulating, and anti-viral agents. <br><br> Description of the Related Art <br><br> The biology of cell proliferation and cell death (apoptosis) is extremely complex, involving multiple intracellular 20 signaling pathways and multiple interacting gene products. Cancer cells may exhibit multiple defects in normal regulatory controls of cell proliferation which allow them to increase in number. Furthermore, cancer cells exhibit defects in mechanisms that are involved in eliminating abnormal cells by multi-step processes 25 referred to as programmed cell death or apoptosis. Thus, combinations of unregulated cell proliferation and suppression of death inducing signaling pathways give cancer cells both growth and survival advantages. <br><br> WO 01/58889 PCT/USO1/04168 <br><br> Whether a cell increases in numbers or not depends on a balance of expression of negatively-acting and positively-acting growth regulatory gene products, and the presence or absence of functional cell death signaling pathways. Negative-acting growth 5 regulatory genes contribute to blockage of cells in the cell cycle. Positive-acting growth regulatory genes stimulate cells to progress through the cell cycle. Genes involved in apoptosis can be either proapoptotic or antiapoptotic, and the dynamic balance between them determines whether a cell lives or dies. <br><br> 10 Cancer cells, in order to survive and increase their numbers, undergo a series of mutational events over time that remove regulatory controls that give them the ability to grow unchecked and survive even in the presence of proapoptotic signals, and develop attributes that permit them to escape detection and 15 removal by the immune response defense system. Cancers may cause death of individuals unless removed by surgery or effectively treated with drugs. <br><br> A wide variety of pathological cell proliferative conditions exist for which novel therapeutic strategies and agents 20 are needed to provide therapeutic benefits. These pathological conditions may occur in almost all cell types capable of abnormal cell proliferation or abnormal responsiveness to cell death signals. Among the cell types that exhibit pathological or abnormal growth and death characteristics are (1) fibroblasts, (2) vascular 25 endothelial cells, and (3) epithelial cells. Thus, novel methods are needed to treat local or disseminated pathological conditions in all or almost all organ and tissue systems of individuals. <br><br> Most cancers, whether they be male specific such as prostate or testicular, or female specific such as breast, ovarian or <br><br> 2 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> cervical or whether they affect males and females equally such as liver, skin or lung, with time undergo increased genetic lesions and epigenetic events, and eventually become highly metastatic and difficult to treat. Surgical removal of localized cancers has proven 5 effective only when the cancer has not spread beyond the primary lesion. Once the cancer has spread to other tissues and organs, the surgical procedures must be supplemented with other more specific procedures to eradicate the diseased or malignant cells. Most of the commonly utilized supplementary procedures for treating diseased 10 or malignant cells such as chemotherapy or bioradiation are not localized to the tumor cells and, although they have a proportionally greater destructive effect on malignant cells, often affect normal cells to some extent. <br><br> Some derivatives of tocopherols, tocotrienols and 15 vitamin E have been used as proapoptotic and DNA synthesis inhibiting agents. Structurally, vitamin E is composed of a chromanol head and an alkyl side chain. There are eight major naturally occurring forms of vitamin E: alpha (a), beta (P), gamma (y), and delta (8) tocopherols and a, p, y, and 8 tocotrienols. 20 Tocopherols differ from tocotrienols in that they have a saturated phytyl side chain rather than an unsaturated isoprenyl side chain. The four forms of tocopherols and tocotrienols differ in the number of methyl groups on the chromanol head (a has three, P and y have two and 8 has one). <br><br> 25 RRR-oc-tocopheryl succinate is a derivative of RRR-a- <br><br> tocopherol that has been structurally modified via an ester linkage to contain a succinyl moiety instead of a hydroxyl moiety at the 6-position of the chroman head. This ester linked succinate moiety of RRR-a-tocopherol has been the most potent form of vitamin E <br><br> 3 <br><br> affecting the biological actions of triggering apoptosis and inhibiting DNA synthesis. This form of vitamin E induces tumor cells to undergo apoptosis, while having no apoptotic inducing effects on normal cells. The major advantage of this form of vitamin E as an anticancer agent is that many cancer cells either express low levels of esterases or do not express esterases that can cleave the succinate moiety, thereby converting the succinate form of RRR-a-tocopherol to the free RRR-a-tocopherol. RRR-a-tocopherol exhibits neither potent antiproliferative nor apoptotic triggering biological activity. However, the ester-linked vitamin E succinate is ineffective in vivo since natural esterases in the host cleave off the succinate moiety, rendering an ineffective anticancer agent, RRR-a-tocopherol. <br><br> The prior art is deficient in the lack of effective means of inhibiting undesirable or uncontrollable cell proliferation in a wide variety of pathophysiological conditions while having no to little effect on normal cells. The present invention fulfills this longstanding need and desire in the art. <br><br> SUMMARY OF THE INVENTION <br><br> In one embodiment of the present invention, <br><br> there is provided a compound having a structural formula wherein X is oxygen, nitrogen or sulfur; Y is oxygen or NR6; <br><br> R1 is R7 or R9; <br><br> 4 <br><br> INTELLECTUAL PROPERTY OFFICE OF N2 <br><br> 23 MAR 2004 RECEIVED <br><br> R2, R3 are independently hydrogen, methyl, -CH2(C6H4)Ci_ 4alkylene-COOH, -C1.4alkylene-COO-CH2 (C6H5), -Ci.4alkylene-CO-NH-CH2(C6H5), saccharide, -Ci.4alkylene-CH2-NH(2-n)(Ci-4alkyl)n wherein n is 2 or 1; <br><br> R4 is methyl, -CH2(C6H4)Ci.4alkylene-COOH, -C^alkylene-COO-CH2(C6H5), -Ci.4alkylene-CO-NH-CH2(C6H5), saccharide, -Q. 4alkylene-CH2-NH(2.n)(Ci.4alkyl)n wherein n is 2 or 1; <br><br> R5 is methyl or R8; <br><br> R6 is hydrogen or methyl <br><br> R7 is -Cx-ioalkylene-COOH, -Ci_4alkylene-CONH2, -Q. 4alkylene-COO-C1.4alkyl, -Q _4alkylene-C0N(Cx _4alkylene-COOH)2, -Q_ 4alkylene-OH, -Cx.4all&lt;ylene-NH3-halo or -C1.4alkylene-0S02NH(C1. 4alkyl); <br><br> R8 is -C7.i7alkyl, -COOH, -C7.17 olefinic group containing 3 to 5 ethylenic bonds, -C=C-C00-Ci-4alkyl, or -Ci.4alkylene-C00-Ci-4 alkyl; <br><br> R9 is -Ci^alkylene-O-Cj.4alkyl, -Ci-i0alkylene-CO-SH, -Ci. 4alkylene-CO-S(Ci-4alkyl), -Ci-4alkylene-CS-NH2, saccharide, alkoxy-linked saccharide, -C1.4alkylene-CO-NH(2.n)(Ci^alkyl)n wherein n is 2 or 1, -C1.4alkylene-S02-0(Ci-4alkyl), -Ci.4alkylene-0S02-0(Ci.4alkyl), -Ci.4alkylene-0P(0-C1.4all^l)3, or -Ci-i0alkylene-CN; and with the proviso that X and Y cannot both be oxygen when <br><br> R1 is R7, <br><br> R2, R3 are independently hydrogen or Ci.4alkyl; R4 is Ci.4alkyl; and R5 is R8. <br><br> In another embodiment, there is provided a pharmaceutical composition comprising a compound of the invention and a pharmaceutically acceptable carrier. <br><br> In a further embodiment, there is provided a use of a compound of the invention in the manufacture of a medicament to treat a cell proliferative disease. <br><br> INTELLECTUAL PROPERTY OFFICE OF H2 <br><br> 2 3 MAR 200^ <br><br> RECEIVED <br><br> Described herein is a compound having a structural formula <br><br> RV <br><br> n wherein X is oxygen, nitrogen or sulfur; Y is selected from the group consisting of oxygen, nitrogen and sulfur wherein when Yis oxygen or nitrogen, n is 1 and when Yis sulfur, n is 0; R1 is selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, heteroaryl, carboxylic acid, carboxylate, carboxamide, ester, thioamide, thiolester, thiolacid, saccharide, alkoxy-linked saccharide, amine, sulfonate, sulfate, phosphate, alcohol, ether and nitrile; R2 is selected from the group consisting of hydrogen, methyl, benzyl carboxylic acid, benzyl carboxylate, benzyl carboxamide, benzyl ester, saccharide and amine; R3 is selected from the group consisting of hydrogen, methyl, benzyl carboxylic acid, benzyl carboxylate, benzyl carboxamide, benzylester, saccharide and amine; R4 is selected from the group consisting of methyl, benzyl carboxylic acid, benzyl carboxylate, benzyl carboxamide, benzylester, saccharide and amine; and R5 is selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, heteroaryl, carboxyl, amide and ester; wherein when Y is nitrogen, said nitrogen is substituted with R6, wherein R6 is selected from the group consisting of hydrogen and methyl; wherein when X is oxygen, R2 is methyl, R3 is methyl, R4 is methyl and R5 is phytyl, R1 is not butyric acid. <br><br> a cell proliferative disease comprising administering to an animal a pharmacologically effective dose of a compound having a structural formula <br><br> Also described is a method for the treatment of i4 <br><br> INTELLECTUAL PROPERTY OFRCE OF N.Z <br><br> 23 MAR 2004 <br><br> 5 <br><br> RECEIVED <br><br> wherein X is oxygen, nitrogen or sulfur; Y is selected from the group consisting of oxygen, nitrogen and sulfur wherein when Yis oxygen or nitrogen, n is 1 and when Yis sulfur, n is 0; R1 is selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, heteroaryl, 5 carboxylic acid, carboxylate, carboxamide, ester, thioamide, thiolester, thiolacid, saccharide, alkoxy-linked saccharide, amine, sulfonate, sulfate, phosphate, alcohol, ether and nitrile; R2 is selected from the group consisting of hydrogen, methyl, benzyl carboxylic acid, benzyl carboxylate, benzyl carboxamide, benzyl 10 ester, saccharide and amine; R3 is selected from the group consisting of hydrogen, methyl, benzyl carboxylic acid, benzyl carboxylate, benzyl carboxamide, benzylester, saccharide and amine; R4 is selected from the group consisting of methyl, benzyl carboxylic acid, benzyl carboxylate, benzyl carboxamide, 15 benzylester, saccharide and amine; and R5 is selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, heteroaryl, carboxyl, amide and ester; wherein when Y is nitrogen, said nitrogen is substituted with R6, wherein R6 is selected from the group consisting of hydrogen and methyl.. <br><br> 20 Also described is a pharmaceutical composition comprising a compound disclosed herein and a pharmaceutically acceptable carrier. <br><br> Also described is a method of inducing apoptosis of a cell, comprising the step of contacting said cell with a pharmacologically effective does of a compound of the present invention. <br><br> Other and further aspects, features, benefits, and advantages of the present invention will be apparent from the <br><br> INTELLECTUAL PROPERTY OFFICE OF N.Z <br><br> 23 MAR 2004 <br><br> RECEIVED <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> following description of the presently preferred embodiments of the invention given for the purpose of disclosure. <br><br> 5 BRIEF DESCRIPTION OF THE DRAWINGS <br><br> So that the matter in which the above-recited features, advantages and objects of the invention, as well as others which will become clear, are attained and can be understood in detail, more ^10 particular descriptions of the invention are briefly summarized. <br><br> above may be had by reference to certain embodiments thereof which are illustrated in the appended drawings. These drawings form a part of the specification. It is to be noted; however, that the appended drawings illustrate preferred embodiments of the 15 invention and therefore are not to be considered limiting in their scope. <br><br> Figure 1 shows general structure of tocopherol, tocotrienol and other chroman-based compounds. <br><br> Figure 2 shows general tocopherol-based compounds 20 1-29 presently synthesized and tested. <br><br> Figures 3A, 3B and 3C shows general synthetic organic approaches for the chemical variation of chromanol compounds at position R1. <br><br> Figure 4 shows general synthetic organic approaches 25 for the chemical variation of chromanol compounds at position R2. <br><br> Figure 5 shows general synthetic organic approaches for the chemical variation of chromanol compounds at position R3 and R4. <br><br> 7 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> Figure 6 shows general synthetic organic approaches for the chemical variation of chromanol compounds at position R5. <br><br> Figure 7 shows general synthetic organic approaches for the all-racemic 1-aza-a-tocopherol analogs. Figure 7 A shows 5 the synthetic scheme for compounds 31-38 and Figure 7B shows the synthetic scheme for compounds 39-43. <br><br> Figure 8 shows the mean body weights of non-tumor Balb/c female mice at day 11 and day 23 of a maximum tolerated dose study. <br><br> 10 Figure 9 shows the comparison of percent mean tumor weight following treatments in MDA MB-435 Human Breast Cancer Cells (Figure 9A), DU-145 Human Prostrate Cancer Cells (Figure 9B) and HT-29 Human Colon Cancer Cells (Figure 9C). <br><br> 15 <br><br> DETAILED DESCRIPTION OF THE INVENTION <br><br> The following definitions are given for the purpose of facilitating understanding of the inventions disclosed herein. Any 20 terms not specifically defined should be interpreted according to the common meaning of the term in the art. <br><br> As used herein, the term "individual" shall refer to animals and humans. <br><br> As used herein, the term "biologically inhibiting" or 25 "inhibition" of the growth of proliferating cells shall include partial or total growth inhibition and also is meant to include decreases in the rate of proliferation or growth of the cells. The biologically inhibitory dose of the composition of the present invention may b e determined by assessing the effects of the test element on target <br><br> 8 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> malignant or abnormally proliferating cell growth in tissue culture, tumor growth in animals and cell culture or any other method known to those of ordinary skill in the art. <br><br> As used herein, the term "induction of programmed cell 5 death or apoptosis" shall include partial or total cell death with cells exhibiting established morphological and biochemical apoptotic characteristics. The dose of the composition of the present invention that induces apoptosis may be determined by assessing the effects of the test element on target malignant or abnormally 10 proliferating cell growth in tissue culture, tumor growth in animals and cell culture or any other method known to those of ordinary skill in the art. <br><br> As used herein, the term "induction of cell cycle arrest" shall include growth arrest due to treated cells being blocked in 15 GO/G1 or G2/M cell cycle phase. The dose of the composition of the present invention that induces cell cycle arrest may be determined by assessing the effects of the test element on target malignant or abnormally proliferating cell growth in tissue culture, tumor growth in animals and cell culture or any other method 20 known to those of ordinary skill in the art. <br><br> As used herein, the term "induction of cellular differentiation" shall include growth arrest due to treated cells being induced to undergo cellular differentiation, a stage in which cellular proliferation does not occur. The dose of the composition 25 of the present invention that induces cellular differentiation may be determined by assessing the effects of the test element on target malignant or abnormally proliferating cell growth in tissue culture, tumor growth in animals and cell culture or any other method known to those of ordinary skill in the art. <br><br> 9 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> The' present invention provides tocopherols, tocotrienols, and other chroman derivatives with or without derivatives of saturated phytyl or unsaturated isoprenyl side chains and analogs; e.g., azo- and thiol- analogs, thereof. Utilizing ethers 5 and several other chemical linkages to attach different moieties to tocopherol, tocotrienol and other chroman derivatives, novel anticancer compounds, for in vivo use, are produced. The general structures of the novel compounds of the present invention are shown in Figure 1, the preferred compounds are listed in Figure 2 10 and possible routes for their syntheses are provided in Figures 3-7. The novel features of these molecules include chemical funtionalization of positions R1 - R5 of the chroman structure, and chemical functionalization of the phytyl and isoprenyl side chains, particularly compounds based on tocopherols and tocotrienols 15 (Figure 1). Additionally, compounds with heteroatom substitutions (N or S) for the chroman ring oxygen are presented (Figure 7). Particularly preferred compounds include 2,5,7,8-tetramethyl-(2R-(4R,8R,12-trimethyltridecyl)chroman-6-yloxy)acetic acid (1), 2,5,7,8-tetramethyl-(2R-(4R,8R,12-trimethyltridecyl)chroman-6-20 yloxy)propionic acid (2), 2,5,8-trimethyl-(2R-(4R,8R,12-trimethyltridecyl)chroman-6-yloxy)acetic acid (7), 2,7,8-trimethyl-(2R-(4R,8R,12-trimethyltridecyl)chroman-6-yloxy)acetic acid (8), 2,8-dimethyl-(2R-(4R,8R,12-trimethyltridecyl) chroman-6-yloxy) acetic acid (9), 2-(N,N-(carboxymethyl)-2(2,5,7,8-tetramethyl-(2R-25 (4R,8R,12-trimethyltridecyl) chroman-6-yloxy) acetic acid (12), 2,5,7,8-tetramethyl-(2RS-(4RS ,8RS, 12-trimethyltridecyl)chrOman-6-yloxy)acetic acid (15), 2,5,7,8-tetramethyl-2R-(2RS,6RS,10-trimethylundecyl)chroman-6-yloxy)acetic acid (17), 3-(2,5,7,8-tetramethyl-(2R-(4R,8,12-trimethyltridecyl)chroman-6- <br><br> 10 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> yloxy)propyl-l-ammonium chloride (19), 2,5,7,8-tetramethyl-(2R-(4r,8R,12-trimethyltridecyl)chroman-3-ene-6-yloxy) acetic acid (20), 2-(2,5,7,8-tetramethyl-(2R-(4R,8,12-trimethyltridecyl) <br><br> chroman-6-yloxy)triethylammonium sulfate (21), 6-( 2,5,7,8-5 tetramethyl-(2R-(4R,8,12-trimethyltridecyl)chroman)acetic acid (22), 2,5,7,8,-tetramethyl-(2R-(heptadecyl)chroman-6-yloxy) <br><br> acetic acid (25), 2,5,7,8,-tetramethyl-2R-(4,8,-dimethyl-l,3,7 EZ nonotrien)chroman-6-yloxy) acetic acid (26), E,Z,RSJRS-(phytyltrimethylbenzenethiol-6-yloxy)acetic acid (27), 1-aza-a-10 tocopherol-6-yloxyl-acetic acid (39), l-aza-oc-tocopherol-6-yloxyl-methyl acetate (40), l-aza-N-methyl-a-tocopherol-6-yloxyl-methyl acetate (41), l-aza-N-methyl-a-tocopherol-6-yloxyl- acetic acid (42), 6-(2,4-Dinitrophenylazo(2,5,7,8-tetramethyl-2-(4,8,12- <br><br> trimethyltridecyl)-l,2,3,4-tetrahydroquinoline (43). 15 The pharmacodynamically designed compounds of the present invention have an improved therapeutic index and are potent inhibitors of cancer cell growth; i.e., they demonstrate high antitumor activity with minimal side effects. These compounds, which can not be readily degraded since there are no known 20 etherases in mammals, may be used in the treatment of cancers and disorders involving excess cell proliferation, as well as for cells that accumulate in numbers due to suppressed cell killing mechanisms, with minimal side effects. The compounds of the present invention inhibit cancer cell growth by induction of apoptosis and DNA 25 synthesis arrest. Induction of apoptosis by these compounds is mediated by activation of the TGF-p, stress kinase, and Fas/Fas ligand signaling pathways. Induction of apoptosis by other pathways, for example, ceramide production, are not excluded. These growth inhibitory properties allow these compounds to be <br><br> 11 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> used in the treatment of proliferative diseases, including cancers of different cell types and lineages, non-neoplastic hyperproliferative diseases, and disorders with defects in apoptotic signaling pathways. Several of the compounds of the present invention are both strong 5 inducers of apoptosis and strong inhibitors of DNA synthesis arrest of tumor cells representing different cellular lineages. <br><br> The therapeutic use of the compounds of the present invention in treatment of cancers and other diseases and disorders involving excess cell proliferation or failure of cells to die is 10 illustrated. The novel derivatives (Tables 1 and 2) were shown at EC50 concentrations to induce apoptosis of human breast cancer cells (MDA MB 435, MDA MB 231, and MCF-7 breast cancer cells), human prostate cancer cells (PC-3, DU-145 and LnCaP), human ovarian tumor cells (C-170), human cervical tumor cells (ME-180), 15 human endometrial cells (RL-95-2), human lymphoid cells (myeloma, Raji, Ramos, Jurkat, and HL-60), colon cancer cells (HT-29 and DLD-1) and lung cancer cells (A-549). The novel derivatives were shown to not induce apoptosis of normal human mammary epithelial cells (HMECs) and immortalized but non-tumorigenic 20 MCF-10A mammary cells. <br><br> These novel compounds and methods of the present invention may be used to treat neoplastic diseases and nonneoplastic diseases. Representative examples of neoplastic diseases are ovarian cancer, cervical cancer, endometrial cancer, bladder 25 cancer, lung cancer, cervical cancer, breast cancer, prostate cancer, testicular cancer, gliomas, fibrosarcomas, retinoblastomas, melanomas, soft tissue sarcomas, osteosarcomas, colon cancer, carcinoma of the kidney, pancreatic cancer, basal cell carcinoma, and squamous cell carcinoma. Representative examples of non- <br><br> 12 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> neoplastic diseases are selected from the group consisting of psoriasis, benign proliferative skin diseases, ichthyosis, papilloma, restinosis, scleroderma and hemangioma. <br><br> The compounds and methods of the present invention 5 may be used to treat non-neoplastic diseases that develop due to failure of selected cells to undergo normal programmed cell death or apoptosis. Representative examples of diseases and disorders that occur due to the failure of cells to die are autoimmune diseases. Autoimmune diseases are characterized by immune cell 10 destruction of self cells, tissues and organs. A representative group of autoimmune diseases includes autoimmune thyroiditis, multiple sclerosis, myasthenia gravis, systemic lupus erythematosus, dermatitis herpetiformis, celiac disease, and rheumatoid arthritis. This invention is not limited to autoimmunity, but includes all 15 disorders having an immune component, such as the inflammatory process involved in cardiovascular plaque formation, or ultra violet radiation induced skin damage. <br><br> The compounds and methods of the present invention may be used to treat disorders and diseases that develop due to 20 virus infections. Representative examples of diseases and disorders that occur due to virus infections are human immunodeficiency viruses (HIV). Since these compounds are working on intracellular signaling networks, they have the capacity to impact on any type of external cellular signal such as cytokines, viruses, bacteria, toxins, 25 heavy metals, etc. <br><br> The methods of the present invention may be used to treat any animal. Most preferably, the methods of the present invention are useful in humans. <br><br> 13 <br><br> WO 01/58889 PCT/US01/04168 <br><br> Generally, to achieve pharmacologically efficacious cell killing and anti-proliferative effects, these compounds and analogs thereof may be administered in any therapeutically effective dose. Preferably, the structurally modified tocopherols and tocotrienols 5 and analogs are administered in a dose of from about 0.1 mg/kg to about 100 mg/kg. More preferably, the structurally modified tocopherols and tocotrienols and analogs are administered in a dose of from about 1 mg/kg to about 10 mg/kg. <br><br> Administration of the compositions of the present 10 invention may be by topical, intraocular, parenteral, oral, intranasal, intravenous, intramuscular, subcutaneous, or any other suitable means. The dosage administered is dependent upon the age, clinical stage and extent of the disease or genetic predisposition of the individual, location, weight, kind of concurrent treatment, if 15 any, and nature of the pathological or malignant condition. The effective delivery system useful in the method of the present invention may be employed in such forms as capsules, tablets, liquid solutions, suspensions, or elixirs, for oral administration, or sterile, liquid forms such as solutions, suspensions or emulsions. For 20 topical use it may be employed in such forms as ointments, creams or sprays. Any inert carrier is preferably used in combination with suitable solubilizing agents, such as saline, or phosphate-buffered saline, or any such carrier in which the compounds used in the method, such as ethanol, acetone, or DMSO, of the present 25 invention have suitable solubility properties. <br><br> There are a wide variety of pathological cancerous and noncancerous cell proliferative conditions and cell accumulations due to absence of normal cellular death for which the compositions and methods of the present invention will provide therapeutic <br><br> 14 <br><br> WO 01/58889 PCT/US01/04168 <br><br> benefits. These pathological conditions may occur in almost all cell types capable of abnormal cell proliferation or defective in programmed cell death mechanisms. Among the cell types which exhibit pathological or abnormal growth or abnormal death are (1) 5 fibroblasts, (2) vascular endothelial cells and (3) epithelial cells. It can be seen from the above that the methods of the present invention is useful in treating local or disseminated pathological conditions in all or almost all organ and tissue systems of individuals. <br><br> 10 It is specifically contemplated that pharmaceutical compositions may be prepared using the novel chroman-based compounds and derivatives and analogs thereof of the present invention. In such a case, the pharmaceutical composition comprises the novel compounds of the present invention and a 15 pharmaceutical^ acceptable carrier. A person having ordinary skill in this art would readily be able to determine, without undue experimentation, the appropriate dosages and routes of administration of the compounds and analogs of the present invention. <br><br> 20 Thus the present invention is directed toward the design and effective use of novel agents that can specifically target cancer cells and either down-regulate growth stimulatory signals, up-regulate growth inhibitory signals, down-regulate survival signals and/or up-regulate death signals. More specifically, this invention 25 creates and characterizes novel agents that activate growth inhibitory factors, trigger death signaling pathways, and inhibit DNA synthesis. <br><br> 15 <br><br> WO 01/58889 PCT/US01/04168 <br><br> The following examples are given for the purpose of illustrating various embodiments of the invention and are not meant to limit the present invention in any fashion. <br><br> 5 EXAMPLE 1 <br><br> Synthetic Organic, Methodology <br><br> The synthesis of a variety of tocopherol, tocotrienol, and other chroman derivatives with or without derivatives of saturated 10 phytyl or unsaturated isoprenyl side chains or analogs thereof is possible via structural modification of the chroman ring system (Figures 3-6) and heteroatom substitutions (N or S) for the chroman ring oxygen (Figure 7A and 7B). The structural variables R1, R2, R3, R4, R5, and X illustrate the groups on the chroman group that are 15 modified and Y represents either oxygen, or heteroatom substitutions (N or S) for the chroman ring oxygen. Using alkylation chemistry, a large number of compounds containing different R1 groups can be synthesized, particularly when X is oxygen. After alkylation, further chemical modification of the R1 20 groups permits the synthesis of a wide range of novel compounds. Bromination of the benzylic methyl groups of the chroman group provide intermediates that permit variation of the R2, R3 and R4 groups. Variation of group R5 is also possible, particularly when starting from the commercially available 6-hydroxy-2,5,7,8-25 tetramethylchroman-2-carboxylic acid. When a heteroatom substitution of nitrogen for the chroman ring oxygen occurs, than nitrogen may be substituted with R6 which is hydrogen or methyl. Variation of Xto groups other than oxygen, which is the identity of X in tocopherols and tocotrienols, can be accomplished using <br><br> 16 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> palladium chemistry (for X = CH2) and nucleophilic aromatic substitution (for X = N or S). Other possible modifications to the chroman structure include unsaturation at the 3-4 positions and ring contraction to produce a five-membered furanyl ring. <br><br> 5 Reagents employed were either commercially available or prepared according to a known procedure. Anhydrous CH2C12 and THF were obtained by distillation. All other solvents used were reagent. Anhydrous reaction conditions were maintained under a slightly positive argon atmosphere in oven-dried glassware. Silica 10 gel chromatography was performed using 230-400 mesh silica purchased from EM Science. Routine 1H- and 13C-NMR spectra were obtained on a Varian Unity spectrometer at 300.132 MHz and 75.033 MHz frequencies, respectively. NMR spectra were referenced to TMS (0 ppm) or to the isotopic impurity peak of 15 CDC13 (7.26 and 77.0 ppm for 'H and 13C, respectively). High resolution electron impact ionization mass spectroscopy was performed by the Mass Spectrometry Center at The University of Texas at Austin. <br><br> 20 EXAMPLE 2 <br><br> Synthesis and Characterization of Novp.1 Tncnphp.rnl Compounds 2.5.7,8-tfttramftthy1-(2R-(4Rr8RT12-tximet.hyHridecy1) chroman-6-yloxy) acetic, acid (1) <br><br> 25 <br><br> 17 <br><br> WO 01/58889 PCT/US01/04168 <br><br> A solution of R,R,R-a-tocopherol (0.5 g, 1.16 mmol) in N,N-dimethylformamide (20 mL) was treated with methyl bromoacetate (3.4 g, 8.3 mmol) and an excess of powdered NaOH (1.2 g, 30 mmol). The resulting yellow slurry was stirred vigorously 5 for 24 h at room temperature. The reaction was acidified with 5 N HC1 and extracted with diethyl ether (3 x 30 ml). The combined ether layers were washed with H20 (3 x 30 ml) and brine (1 x 3 0 ml), and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel 10 chromatography eluting with 19% (v/v) EtOAc and 2% acetic acid in hexanes. The resulting yellow liquid was dissolved in diethyl ether (30 ml), washed with H20 (3 x 20 mL) and brine (1 x 20 mL), and then dried with Na2S04. The resulting solution was concentrated to a light yellow oil and dried in vacuo for 48 h. This yielded 1 as a 15 waxy, off-white solid (0.50 g, 88%). 'H-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-,12'-CH, l'-,2'-,3'-,5'-,6'-,7'-,9'-,10'-,ll'-CH2, 2a-CH3), 1.81 (m, 2H, 3-CH2), 2.07, 2.14, 2.16 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.59 (t, J = 6.6 Hz, 2H, 4-CH2), 4.34 (s, 2H, OCH2); 13C-NMR (CDC13, ppm): 11.7, 20 11.8, 12.7 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.8 (CH2), 28.0 (CH), 31.2 (3-CH2), 32.7, 32.8 (CH), 37.3, 37.4, 37.5, 39.4, 40.0 (CH2), 69.2 (OCH2), 75.0 (2-C), 117.8, 123.2, 125.4, 127.3 (aryl C), 147.0, 148.5 (aryl C-O), 173.7 (COOH); HRMS (CI, m/z): 489.394374 (M + 25 H+, Calc. for C3iH5304 489.394386). All assignments were confirmed using HMQC, DEPT-135, and ^-NOSEY. <br><br> 18 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> 2 T 5 T 7:8 - tetr a rn eth y 1 - (2R - (4R. 8R T11 -tri m e th v 1 tri decv 1) chrom an -6-yloxv) propionic acid (2) <br><br> The compounds 2-6 are synthesized in a manner identical to the synthesis of 1 using the appropriate bromoalkaaoic acids. <br><br> (89% yield). TC-NMR (CDC1/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0-1.6 (m, 24H, 4'-, 8'-, 12'-CH, l'-,2'-,3'-,5'-,6'-,7'-,9'-,10'-,ll'-CH2, 2a-CH3), 1.81 (m, 2H, 3-CH2), 2.09, 2.14, 2.19 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.59 (t, J = 6.6 Hz, 2H, 4-CH2), 2.85 (t, J = 6.4 Hz, 2H, CH2COOH), 3.96 (t, J = 6.4 Hz, 2H, OCH2); 13C-NMR (CDC13, ppm): 11.7, 11.8, 12.7 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.8 (CH2), 28.0 (CH), 31.2 (3-CH2), 32.7, 32.8 (CH), 35.1, 37.3, 37.4, 37.5, 39.4, 40.0 (CH2), 67.5 (OCH2), 74.8 (2-C), 117.5, 122.9. 125.8, 127.8 (aryl C), 147.6, 148.0 (aryl C-O), 177.1 (COOH); HRMS (CI, m/z): 503.408610 (M + H+, Calc. for C32H5504 503.410036). <br><br> HY^&lt; <br><br> o <br><br> 9.; 5,7,8-tp.tramp.tby1 -f2R-(4R T 8R T12-tri m ethyl tri de.r.yl ^ r.hroman-6-yloxy) butyric acid (3) <br><br> HYi&lt; <br><br> o <br><br> 19 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> (85% yield). ^-NMR (CDC1/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 26H, 4'-, 8'-, 12'-CH, l'-,2'-,3'-,5'-,6'-,7'-,9'-,10'-,ll'-CH2, 2a-CH3), 1.81 (m, 2H, 3-CH2), 2.14, 2.17, 2.21 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.62 (t, J = 6.6 Hz, 2H, 4-CH2), 2.72 (t, J = 7.2 Hz, 2H, CH2COOH), 3.74 (t, J = 6.1 Hz, 2H, OCH2); 13C-NMR (CDC13, ppm): 11.7, 11.8, 12.7 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.9 (2a-CH3), 24.4, 24.8, 25.3 (CH2), 28.0 (CH), 30.9, 31.2 (3-CH2), 32.7, 32.8 (CH), 37.3, 37.4, 37.5, 39.4, 40.0 (CH2), 71.3 (OCH2), 74.8 (2-C), 117.5, 122.9. 125.7, 127.7 (aryl C), 147.8, 147.9 (aryl C-O), 178.9 (COOH); HRMS (CI, m/z): 516.424374 (M + H+, Calc. for C33H5704 516.424386). <br><br> 7,5,7, R -t ft tr a m p, th y T - 9.R - (4R, R R, 11-tri m ft th yl tri H e.r,y 1) chrr&gt;man-6-ylr&gt;yv) valftric add (4) <br><br> (90% yield). !H-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, 4a*-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 28H, 4'-, 8'-,12'-CH, l'-,2'-^'-^'-^•-J'-^'-aO'-ai'-CHz, 2a-CH3), 1.81 (m, 2H, 3-CH2), 2.09, 2.14, 2.18 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.49 (t, J = 6.8 Hz, 2H, CH2COOH), 2.59 (t, J = 6.6 Hz, 2H, 4-CH2), 3.68 (t, J = 5.5 Hz, 2H, OCH2); 13C-NMR (CDC13, ppm): 11.7, 11.8, 12.7 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0, 21.4 (CH2), 22.6, 22.7 (CH3), 23.8 (2a- <br><br> 20 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> CH3), 24.4, 24.8 (CH2), 28.0 (CH), 30.0 (CH2), 31.2 (3-CH2), 32.7, <br><br> 32.8 (CH), 35.8, 37.3, 37.4, 37.5, 39.4, 40.0 (CH2), 72.2 (OCH2), <br><br> 74.9 (2-C), 117.8, 123.2. 125.4, 127.3 (aryl C), 147.6, 148.3 (aryl C-O), 178.7 (COOH); HRMS (CI, m/z): 530.433514 (M + H+, Calc. for C34H59O4 530.433516). <br><br> 2-5.7I8-tRtrflmethy1-2R-(4R-8T?11 9.-tri methyl tri decvl^ chrr&gt;man-6-y1oxy)hftxanoic ar.id (5) <br><br> (77% yield). ^-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 30H, 4'-, 8'-,12'-CH, l'-,2'-^'-^'-^'-J'-^'-.lO'-ai'-CH^ 2a-CH3), 1.81 (m, 2H, 3-CH2), 2.08, 2.12, 2.16 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.32 (t, J = 6.5 Hz, 2H, CH2COOH), 2.57 (t, J = 6.6 Hz, 2H, 4-£H2), 3.64 (t, J = 5.5 Hz, 2H, OCH2); 13C-NMR (CDCI3, ppm): 11.8, 11.9, 12.7 (5a-, 7a-, 8a-CH3), <br><br> 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.6, 24.8, 25.7 (CH2), 28.0 (CH), 30.0 (CH2), 31.3 (3-CH2), <br><br> 32.7, 32.8 (CH), 34.0, 37.3, 37.3, 37.4, 39.3, 40.0 (CH2), 72.6 (OCH2), 74.7 (2-C), 117.4, 122.7. 125.4, 127.8 (aryl C), 147.6, 148.2 (aryl C-O), 179.6 (COOH); HRMS (CI, m/z): 545.457026 (M + H+, Calc. for C35H6104 545.456986). <br><br> HY^&lt; <br><br> o <br><br> 21 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> 2T5I7,R-tetramethy1-2R-(4'RJRRJ12-trimethy1tridecyl) chrnman-fi-yloyy^octflnnic acid (£) <br><br> H0y4&lt; <br><br> O <br><br> 5 <br><br> (91% yield). 'H-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, <br><br> 4a'-, 8a'-, 12a'-, lS'-CHs), 1.0 - 1.6 (m, 34H, 4'-, 8'-,12'-CH, l'-,2'-0 ,V5'9'2a-CH3), 1.81 (m, 2H, 3-CH2), 2.08, 2.11, 2.16 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.36 (m, 2H, CH2COOH), 2.58 (t, J = 6.6 Hz, 2H, 4-CH2), 3.62 (t, J = 5.5 Hz, 2H, OCH2); 13C-10 NMR (CDC13, ppm): 11.7, 11.8, 12.7 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.6, 24.8, 25.1, 25.7, 26.6 (CH2), 28.0 (CH), 30.0 (CH2), 31.3 (3-CH2), 32.7, 32.8 (CH), 34.0, 37.3, 37.3, 37.4, 39.3, 40.0 (CH2), 72.7 (OCH2), 74.6 (2-C), 117.6, 122.8. 125.5, 127.6 (aryl C), 147.5, 15 148.3 (aryl C-O), 179.4 (COOH); HRMS (CI, m/z): 573.484396 (M + H+, Calc. for C37H6504 573.488286). <br><br> 215.8-trimftt&gt;iy1-(2R-(4R1RR; T 2-trimethyltndecyll chroman-6-y1oxy)acetic acid (2} <br><br> 20 <br><br> h&lt;xtT° <br><br> o <br><br> A solution of R,R,R-oc-tocopherol (75 mg, 0.18 mmol) in N,N-dimethylformamide (2 mL) was treated with methyl <br><br> 22 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> bromoacetate (0.4 g, 2.8 mmol) and an excess of powdered NaOH (0.5 g, 12.5 mmol). The resulting yellow slurry was stirred vigorously for 24 h at room temperature. The reaction was acidified with 5 N HCl and extracted with diethyl ether (3 x 10 ml). <br><br> 5 The combined ether layers were washed with H20 (3 x 10 ml) and brine (1 x 10 ml), and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel chromatography eluting with 19% (v/v) EtOAc and 2% acetic acid in hexanes. The resulting yellow liquid was dissolved in diethyl ether 10 (30 ml), washed with H20 (3 x 10 mL) and brine (1 x 10 mL), and then dried with Na2S04. The resulting solution was concentrated to a light yellow oil and dried in vacuo for 48 h. This yielded 7 as a waxy, off-white solid (80 mg, 97%). ^-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-15 ,12'-CH, l'-^'-Z-^'-^'-J'-^'-JO'-ai'-CHz, 2a-CH3), 1.81 (m, 2H, <br><br> 3-CH2), 2.12, 2.14 (2 x s, 6H, 5a-, 8a-CH3), 2.61 (t, J = 6.6 Hz, 2H, <br><br> 4-CH2), 4.59 (s, 2H, OCH2), 6.53 (s, 1H, aryl CH); 13C-NMR (CDC13, ppm): 11.2, 16.1 (5a-, 8a-CH3), 19.6, 19.7 (CH3), 20.7, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.8 (CH2), 27.9 (CH), 31.2 <br><br> 20 (3-CH2), 32.7, 32.8 (CH), 37.2, 37.4, 37.5, 39.4, 40.0 (CH2), 66.8 (OCH2), 74.8 (2-C), 113.8, 120.7, 123.1, 127.3 (aryl C), 147.1, 148.2 (aryl C-O), 175.3 (COOH); HRMS (CI, m/z): 475.377840 (M + H+, Calc. for C30H5IO4 475.378736). <br><br> 25 <br><br> 23 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> ?. 17,8-tri m p.th yl -(2R - (4R, RR, 1 2-tri m ethyl tri decyl ^ chrnm an-6-vlnyy )a.cetic acid (8^ <br><br> A solution of R,R,R-oc-tocopherol (100 mg, 0.24 mmol) in N,N-dimethylformamide (5 mL) was treated with methyl bromoacetate (1.1 g, 7.4 mmol) and an excess of powdered NaOH (1.0 g, 25 mmol). The resulting yellow slurry was stirred vigorously 10 for 24 h at room temperature. The reaction was acidified with 5 N HCl and extracted with diethyl ether (3 x 10 ml). The combined ether layers were washed with HzO (3 x 10 ml) and brine (1 x 10 ml), and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel 15 chromatography eluting with 19% (v/v) EtOAc and 2% acetic acid in hexanes. The resulting yellow liquid was dissolved in diethyl ether (30 ml), washed with H20 (3 x 10 mL) and brine (1 x 10 mL), and then dried with Na2S04. The resulting solution was concentrated to a light yellow oil and dried in vacuo for 48 h. This yielded 8 as a 20 waxy, off-white solid (110 mg, 97%). 'H-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-, 12-CH, 2a-CH3), 1.81 (m, 2H, <br><br> 3-CH2), 2.12, 2.19 (2 x s, 6H, 7a-, 8a-CH3), 2.61 (t, J = 6.6 Hz, 2H, <br><br> 4-CH2), 4.59 (s, 2H, OCH2), 6.39 (s, 1H, aryl CH); 13C-NMR (CDC13, 25 ppm): 11.9, 12.0 (7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.7, 21.0 (CH2), <br><br> 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.8 (CH2), 27.9 (CH), 31.2 <br><br> 24 <br><br> WO 01/58889 PCT/US01/04168 <br><br> (3-CH2), 32.7, 32.8 (CH), 37.2, 37.4, 37.5, 39.4, 40.0 (CH2), 66.6 (OCH2), 75.7 (2-C), 110.6, 117.7, 125.0, 126.3 (aryl C), 146.9, 148.7 (aryl C-O), 175.0 (COOH); HRMS (CI, m/z): 475.377962 (M + H+, Calc. for C3oH5104 475.378736). <br><br> 2.T 8-di methyl-(9.R-(4R; RR T1 2-tri methyl tridecy1)chroman-6-y1oxy)acetin acid (9) <br><br> 10 A solution of R,R,R-a-tocopherol (100 mg, 0.25 mmol) in <br><br> N,N-dimethylformamide (5 mL) was treated with methyl bromoacetate (1.1 g, 7.4 mmol) and an excess of powdered NaOH (1.0 g, 25 mmol). The resulting yellow slurry was stirred vigorously for 24 h at room temperature. The reaction was acidified with 5 N 15 HCl and extracted with diethyl ether (3 x 10 ml). The combined ether layers were washed with H20 (3 x 10 ml) and brine (1 x 10 ml),and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel chromatography eluting with 19% (v/v) EtOAc and 2% acetic acid in 20 hexanes. The resulting yellow liquid was dissolved in diethyl ether (30 ml), washed with H20 (3 x 10 mL) and brine (1 x 10 mL), and then dried with Na2S04. The resulting solution was concentrated to a light yellow oil and dried in vacuo for 48 h. This yielded 9 as a waxy, off-white solid (111 mg, 98%). ]H-NMR (CDCI3/TMS, ppm): 25 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-, 12-CH, 1'-, 2'-,3'-,5'-,6,-,7'-,9,-,10'-,ir-CH2, 2a-CH3), 1.81 (m, 2H, <br><br> 25 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> 3-CH2), 2.15 (s, 3H, 8a-CH3), 2.71 (t, J = 6.6 Hz, 2H, 4-CH2), 4.59 (s, 2H, OCH2), 6.48 (d, J = 3.0 Hz, 1H, aryl CH), 6.61 (d, J = 3.0 Hz, 1H, aryl CH); 13C-NMR (CDC13, ppm): 16.2 (8a-CH3), 19.6, 19.7 (CH3), 21.0 (CH2), 22.6, 22.7 (CH3), 24.0 (2a-CH3), 24.4, 24.8 (CH2), 27.9 5 (CH), 31.2 (3-CH2), 32.7, 32.8 (CH), 37.2, 37.4, 37.5, 39.4, 40.0 (CH2), 65.7 (OCH2), 75.8 (2-C), 112.3, 115.6, 121.1, 127.5 (aryl C), 147.2, 149.9 (aryl C-O), 174.8 (COOH); HRMS (CI, m/z): 460.3552022 (M + H+, Calc. for C30H5iO4 460.355262). <br><br> 10 7,5,7,8- tetram et.hy1-(?.R-(4R.8R,1 2- tri m et.h v 1 tri d ecv 1) chroman-6-y1oxy)acetamide (10) <br><br> 15 A solution of 1 (0.1 g, 0.2 mmol) in CH2C12 (5 mL) was treated with N-hydroxysuccinimide (26 mg, 0.23 mmol) and dicyclohexylcarbodiimide (46 mg, 0.23 mmol). After 2 min, a white precipitate formed. The resulting suspension was stirred for 2 h. The reaction stirred for an additional 6 h. The reaction mixture was 20 cooled to - 30 °C and filtered. The filtrate was concentrated and the resulting colorless oil was purified by silica gel chromatography eluting with EtOAc (35%, v/v) in hexanes. This yielded a white solid (75 mg, 76%). *H-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8«-, 12'-CH, l,-,2,-,3,-,5,-,6'-25 ,7'-,9'-,10'-,ll'-CH2, 2a-CH3), 1.81 (m, 2H, 3-CH2), 2.10, 2.12, 2.16 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.59 (t, J = 6.6 Hz, 2H, 4-CH2), 4.19 (s, <br><br> 26 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> 2H, OCH2), 6.36, 6.92 (2 x broad, 2H, NH); 13C-NMR (CDC13, ppm): 11.7, 11.8, 12.7 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.8 (CH2), 28.0 (CH), 31.2 (3-CH2), 32.7, 32.8 (CH), 37.3, 37.4, 37.5, 39.4, 40.0 (CH2), 5 70.9 (OCH2), 74.9 (2-C), 117.8, 123.3. 125.4, 127.3 (aryl C), 146.5, 148.4 (aryl C-O), 172.1 (COOH); HRMS (CI, m/z): 488.409341 (M + H+, Calc. for C3lH54N03 488.410370). <br><br> M ethyl 2,5,7,8-tetra m et hy 1 - (2R - (4R, 8 R,! 2-10 trimethyUridecyl) chroman-6-ylnxy^r.p.tate (11 ^ <br><br> A solution of 1 (0.1 g, 0.2 mmol) in CH2C12 (5 mL) was treated with N,N-dimethylaminopyridine (26 mg, 0.23 mmol), 15 methanol (1 ml) and dicyclohexylcarbodiimide (46 mg, 0.23 mmol) After 2 min, a white precipitate formed. The resulting suspension was stirred for 6 h. The reaction mixture was cooled to -30 "C and filtered. The filtrate was concentrated and the resulting colorless oil was purified by silica gel chromatography eluting with EtOAc 20 (40%, v/v) in hexanes. This yielded a white solid (82 mg, 80%). XH-NMR (CDC1/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-,12'-CH, l,-,2,-,3,-,5,-,6'-,7,-,9,-,10'-,l 1'-CH2, 2a-CH3), 1.81 (m, 2H, 3-CH2), 2.10, 2.16, 2.20 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.59 (t, J = 6.6 Hz, 2H, 4-CH2), 3.85 (s, 3H, OCH3), 25 4.32 (s, 2H, OCH2); 13C-NMR (CDC13, ppm): 11.7, 11.8, 12.7 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 <br><br> 27 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> (2a-CH3), 24.4, 24.8 (CH2), 28.0 (CH), 31.2 (3-CH2), 32.7, 32.8 (CH), 37.3, 37.4, 37.5, 39.4, 40.0 (CH2), 50.2 (OCH3), 69.8 (OCH2), 74.9 (2-C), 117.6, 123.0, 125.6, 127.5 (aryl C), 147.6, 148.2 (aryl C-O), 169.8 (COOH); HRMS (CI, m/z): 503.408411 (M + H+, Calc. 5 for C32H5504 503.410036). <br><br> 2-(NTN-(carho-)cymfithy1)-2(2T5T7T8-t.et.rame,t.hy1-(2R-(4RT8R,12-t.rimethyltri decy1)chroman-6-y1oxy)acetic acid (12) <br><br> A solution of 1 (0.2 g, 0.4 mmol) in CH2C12 (5 mL) was treated with diethyl iminodiacetate (77 mg,0.4mmol) and 0-7-azabenzotriazol-l-yl-N,N,N',N'-tetramethyuronium hexafluorophosphate (HATU) (46 mg, 0.23 mmol). After 12 h, the 15 reaction mixture was concentrated to a paste and then purified by silica gel chromatography eluting with EtOAc (30%, v/v) in hexanes. This yielded the desired diester intermediate as colorless oil (150 0 mg, 55%). 'H-NMR (CDC1/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 30H, 4'-, 8'-,12'-CH, l,-,2,-,3,-,5,-,6,-20 ,7'-,9'-,10'-,ll'-CH2, 2a-CH3), 1.78 (m, 2H, 3-CH2), 2.08, 2.13, 2.17 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.58 (t, J = 6.8 Hz, 2H, 4-CH2), 4.19, 4.22 (q, J = 7.4 Hz, 4H, OCH2), 4.30, 4.33, 4.42 (3 x s, 6H, 2 x NCH2, OCH2); 13C-NMR (CDC13, ppm): 11.7, 11.8, 12.7 (5a-, 7a-, 8a-CH3), 14.0 (CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 25 23.8 (2a-CH3), 24.4, 24.8 (CH2), 28.0 (CH), 31.2 (3-CH2), 32.7, 32.8 (CH), 37.3, 37.4, 37.5, 39.4, 40.0 (CH2), 48.1, 49.4 (NCH2), 61.2, <br><br> 28 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> 61.5 OCH2), 71.8 (OCH2), 74.8 (2-C), 117.5, 122.9. 125.6, 127.4 (aryl C), 148.0, 148.1 (aryl C-O), 168.8, 169.0 (CO); MS (CI, m/z): 660 (M + H+, Calc. for C39H65N07 659.47610). <br><br> A solution of the diester intermediate (0.15 g, 0.23 5 mmol) in ethanol (4 ml) was treated with 1 N NaOH (1 ml). The resulting cloudy mixture was stirred at 70 °C for 15 h. The reaction mixture was acidified with 1 N HCl and the ethanol was removed in vacuo. The resulting aqueous solution was extracted with CHC13 (5 x 20 ml) and the combined organic layers dried with Na2S04. This 10 yielded 12 (0.13 g, 52%) as a white solid. !H-NMR (CDC1/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-, 12'-CH, 1'-, 2,-,3'-,5,-,6,-,7'-,9,-,10'-,l l'-CH2, 2a-CH3), 1.70 (m, 2H, 3-CH2), 2.01, 2.05, 2.08 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.47 (m, 2H, 4-CH2), 4.18 (m, 4H, 2 x NCH2), 4.31 (m, 2H, OCH2); 13C-15 NMR (CDC13, ppm): 11.5, 11.6, 12.4 (5a-, 7a-, 8a-CH3), 19.4, 19.5 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.8 (CH2), 28.0 (CH), 31.2 (3-CH2), 32.4, 32.5 (CH), 37.0, 37.2, 37.5, 39.1, 40.0 (CH2), 48.1, 49.4 (NCH2), 71.1 (OCH2), 74.8 (2-C), 117.5, 122.9. 125.4, 127.2 (aryl C), 147.8, 148.1 (aryl C-O), 168.8, 169.0 20 (CO); HRMS (CI, m/z): 604.420882 (M + H-, Calc. for C35H58N07 604.421329). • <br><br> 2-(2J5J7,8-tetramethyl-(2R-(4R.8R, 1 2-trimethyltridecyl) r.hrnman-6-yloyy))ftfhati-1 -ol (13) <br><br> 25 <br><br> H0^0 <br><br> 29 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> A solution of R,R,R-a-tocopherol (0.5 g, 1.16 mmol) in N,N-dimethylformamide (20 mL) was treated with iodoethanol (1.7 g, 10 mmol) and an excess of powdered NaOH (2.5 g, 63 mmol). The resulting yellow slurry was stirred vigorously for 24 h at room 5 temperature. The reaction was acidified with 5 N HCl and extracted with diethyl ether (3 x 30 ml). The combined ether layers were washed with H20 (3 x 30 ml) and brine (1 x 30 ml), and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel chromatography eluting with 30% 10 (v/v) EtOAc and 2% acetic acid in hexanes. The resulting yellow liquid was dissolved in diethyl ether (30 ml), washed with H20 (3 x 20 mL) and brine (1 x 20 mL), and then dried with Na2S04. The resulting solution was concentrated to a light yellow oil and dried in vacuo for 48 h. This yielded 13 as yellow oil (0.40 g, 73%). 1H-15 NMR (CDCI3/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-, 12'-CH, l,-,2'-,3'-,5,-,6,-,7,-,9,-,10,-,l 1'-CH2, 2a-CH3), 1.81 (m, 2H, 3-CH2), 2.07, 2.14, 2.16 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.59 (t, J = 6.6 Hz, 2H, 4-CH2), 3.79 (m, 2H, OCH2), 3.94 (m, 2H, OCH2); 13C-NMR (CDC13, ppm): 11.7, 11.8, 12.7 (5a-, 20 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.8 (CH2), 28.0 (CH), 31.2 (3-CH2), 32.7, 32.8 (CH), 37.3, 37.4, 37.5, 39.4, 40.0 (CH2), 63.1, 69.2 (OCH2), 75.0 (2-C), 117.8, 123.4, 126.4, 128.3 (aryl C), 149.2, 149.5 (aryl C-O); MS (CI, m/z): 475 (M + H+, Calc. for C31H5403 474.40729). <br><br> 25 <br><br> 30 <br><br> WO 01/58889 PCT/US01/04168 2-f2.5T7.8-pent.amathy1r:hroman-6-y1oxy)acettc acid <br><br> (14) <br><br> A solution of 2,2,5,7,8-pentamethyl-6-chromanol (0.3 g, 5 1.36 mmol) in N,N-dimethylformamide (20 mL) was treated with methyl bromoacetate (0.8 g, 5.3 mmol) and an excess of powdered NaOH (0.7 g, 18 mmol). The resulting yellow slurry was stirred vigorously for 24 h at room temperature. The reaction was acidified with 5 N HCl and extracted with diethyl ether (3 x 30 ml). 10 The combined ether layers were washed with H20 (3 x 30 ml) and brine (1 x 30 ml), and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel chromatography eluting with 30% (v/v) EtOAc and 2% acetic acid in hexanes. The resulting yellow liquid was dissolved in diethyl ether 15 (30 ml), washed with H20 (3 x 20 mL) and brine (1 x 20 mL), and then dried with Na2S04. The resulting solution was concentrated to a light yellow oil and dried in vacuo for 48 h. This yielded 14 as a white solid (0.31 g, 82%). ^H-NMR (CDC13/TMS, ppm): 1.31 (s, 6H, CH3), 1.81 (t, J = 7.8 Hz, 3-CH2), 2.10, 2.16, 2.19 (3 x s, 9H, 5a-, 7a-20 , 8a-CH3), 2.61 (t, J = 7.8 Hz, 2H, 4-CH2), 4.39 (s, 2H, OCH2); 13C-NMR (CDC13, ppm): 11.7, 11.8, 12.7 (5a-, 7a-, 8a-CH3), 20.9, 26.8, 32.7 (alkyl), 69.1, (OCH2), 72.9 (2-C), 117.5, 123.2, 125.5, 127.3 (aryl), 147.0, 148.6 (O-aryl), 173.8 (COOH); HRMS (CI, m/z): 279.159238 (M + H+, Calc. for C,6H2304 279.159634). <br><br> 25 <br><br> 3 1 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> 2,57, 8-tp.tram p.thv 1 -(9.R S -(4R S T 8R S, 17-tri m p,th y 1 tri ri er.y 1) chromaTi-6-ylnxy)acetic acid (.15). <br><br> A solution of all racemic -a-tocopherol (0.5 g, 1.16 5 mmol) in N,N-dimethylformamide (20 mL) was treated with methyl bromoacetate (3.4 g, 8.3 mmol) and an excess of powdered NaOH (1.2 g, 30 mmol). The resulting yellow slurry was stirred vigorously for 24 h at room temperature. The reaction was acidified with 5 N HCl and extracted with diethyl ether (3 x 30 ml). The combined 10 ether layers were washed with H20 (3 x 30 ml) and brine (1 x 30 ml),and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel chromatography eluting with 19% (v/v) EtOAc and 2% acetic acid in hexanes. The resulting yellow liquid was dissolved in diethyl ether 15 (30 ml), washed with H20 (3 x 20 mL) and brine (1 x 20 mL), and then dried with Na2S04. The resulting solution was concentrated to a light yellow oil and dried in vacuo for 48 h. This yielded 15 as a waxy, off-white solid (80%). rH-NMR (CDCl/TMS, ppm): 0.88 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-,12'-CH, 1'-20 ^'-^'-.S'-^'-J'-^'-.lO'-ai'-CH^ 2a-CH3), 1.84 (m, 2H, 3-CH2), 2.07, 2.14, 2.16 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.61 (t, J = 6.6 Hz, 2H, 4-CH2), 4.34 (s, 2H, OCH2); 13C-NMR (CDC13, ppm): 11.5, 11.7, 12.6 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.3 (CH2), 22.6, 22.8 (CH3), 23.8 (2a-CH3), 24.5, 24.9 (CH2), 29.0 (CH), 31.6 (3-CH2), 25 32.6, 32.8 (CH), 37.5, 37.8, 37.9, 39.5, 41.0 (CH2), 69.3 (OCH2), 75.1 (2-C), 117.9, 123.3, 125.5, 127.3 (aryl C), 147.0, 148.0 (aryl <br><br> 32 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> C-O), 173.9 (COOH); HRMS (CI, m/z): 489.394375 (M + H+, Calc. for C31H53O4 489.394383). <br><br> 2.517T8-tp.tramp,thy1-r9.R-rcarhnYy^chroman-6-y1oxy^ 5 acp,t.ic acid (16) <br><br> A solution of (-)-(R)-6-hydroxy-2,5,7,8- <br><br> tetramethylchroman-2-carboxylic acid (0.34g, 1.36 mmol) in N,N-10 dimethylformamide (20 mL) was treated with methyl bromoacetate (0.8 g, 5.3 mmol) and an excess of powdered NaOH (0.7 g, 18 mmol). The resulting yellow slurry was stirred vigorously for 24 h at room temperature. The reaction was acidified with 5 N HCl and extracted with diethyl ether (3 x 30 ml). The combined ether layers 15 were washed with H20 (3 x 30 ml) and brine (1 x 30 ml), and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel chromatography eluting with 30% (v/v) EtOAc and 2% acetic acid in hexanes. The resulting yellow liquid was dissolved in diethyl ether (30 ml), washed with H20 (3 x 20 20 mL) and brine (1 x 20 mL), and then dried with Na2S04. The resulting solution was concentrated to light yellow oil and dried in vacuo for 48h. This yielded 16 as a white solid (0.33g, 80%). lH-NMR (CDCI3/TMS, ppm): 1.52 (s, 3H, 2a-CH3), 2.10 (m, 2H, 3-CH2), 2.12, 2.16, 2.19 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.56 (t, J = 6.5 Hz, 2H, 25 4-CH2), 4.36 (s, 2H, OCH2). <br><br> 33 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> 215T7T8-tetramftthy1-2R-(9.RS1fiR.Si 1 0-trimpfhylnndp.r.yl^ cfirnma.n-6-ylnyy)acftt.tc acid (17^ <br><br> 5 A solution of lOg (40mmol) of (-)-(S)-6-hydroxy- <br><br> 2,5,7,8-tetramethylchroman-2-carboxylic acid and 0.5g of p-toluenesulfonic acid monohydrate in 200 ml of methanol was stirred and refluxed for 4hr. After cooling, the solution was diluted with water and extracted with diethyl ether. The combined ether 10 layers were washed with saturated aqueous sodium bicarbonate solution , H20 , and brine (1 x 30 ml), and then dried with Na2S04. The resulting solution was concentrated and dried in vacuo for 48 h. This yielded 10 g (95%) of methyl (-)-(S)- 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylate as a colorless solid which was 15 used without further purification. ^-NMR (CDC13/TMS, ppm): 1.52 (s, 3H, 2a-CH3), 2.10 (m, 2H, 3-CH2), 2.12, 2.16, 2.19 (3 x s, 9H, 5 a-, 7a-, 8a-CH3), 2.56 (t, J = 6.5 Hz, 2H, 4-CH2), 3.55 (s, 3H, OCH3); MS (CI, m/z): 264.422 M + H+, Calc. for ClsH20O4 265.3224. <br><br> To a solution of 2g (7.58mmol) of this ester in 7.5ml of 20 N, N-dimethylformamide (DMF) was added 2.6 g (18.8mmol) of anhydrous granular potassium carbonate followed by 2.3 ml ( 20 mmol) of benzylchloride. The resulting slurry was stirred at RT for 41 h then poured into 50 ml of water and worked up with ether in the usual way. The product was freed of excess benzyl chloride at 25 50° under high vacuum. There was obtained 2.69g (100%) of pure (TLC) (-)-(S)-6-benzyloxy-2,5,7,8-tetramethyl-chroman-2carboxylic <br><br> 34 <br><br> WO 01/58889 PCT/USO1/04168 <br><br> acid methyl ester as a yellow solid, m.p. 102-106°. The analytical specimen of this compound was prepared as a colorless solid m.p. 108-109° (from ether/methanol). 'H-NMR (CDCl/TMS, ppm): 1.54 (s, 3H, 2a-CH3), 2.01 (m, 2H, 3-CH2), 2.14, 2.17, 2.19 (3 x s, 9H, 5a-5 , 7a-, 8a-CH3), 2.51 (t, J = 6.7 Hz, 2H, 4-CH2), 3.64 (s, 3H, OCH3), 5.12(s, 2H, 6-OCH2), 7.15 (m,5H, ArH); MS (CI, m/z): 355.232 M + H+, Calc. for C22H2504 354.448. <br><br> A solution of 3.54g (lOmmol) of the above ether ester, in 20 ml of toluene and 10ml of CH2C12 was stirred with cooling 10 from dry ice/acetone bath while 12 ml (18 mmol) of 25% disobutylaluminum hydride in toluene (Texas Alkyls) was added dropwise, over 10 min. After stirring at ca. -70° for 30 min, the reaction mixture was cautiously decomposed (-70°) with 10 ml of MeOH. Following the addition of 50 ml of water and 50 ml of IN 15 aqueous H2S04 solution, the mixture was warmed to RT, and worked up with ether in the usual way giving 3.2 g (100%) of crude aldehyde ((+) S-6-Benzyloxy-2,5,7,8-tetramethylchroman-2-carbaldhyde) as a viscous oil which was purified by silica gel chromatography eluting with 19% (v/v) EtOAc in hexane. XH-NMR 20 (CDCI3/TMS, ppm) : 1.53 (s, 3H, 2a-CH3), 2.11 (m, 2H, 3-CH2), 2.24, 2.27, 2.29 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.481 (t, J = 6.7 Hz, 2H, 4-CH2), 5.19(s, 2H, 6-OCH2), 7.20 (m,5H, ArH), 9.6(s,lH, CHO); MS (CI, m/z): 325.332 M + H+, Calc. for C2iH2403 324.422. <br><br> A solution of 9.6g of pseudoionone was dissolved in 10 0 25 ml of 95% ethanol; after 0.68 g of sodium borohydride in ethanol had been added at room temperature, the mixture was stirred for 2 hr and then left standing overnight. The mixture was added to a solution of 2 g of sodium hydroxide in 500 ml of water. The mixture was extracted with ether, and the ether extract was washed <br><br> 35 <br><br> WO 01/58889 PCT/US01/04168 <br><br> with water, dried, and concentrated. The distillation of the residual oil in vacuo gave a colorless oil (pseudoionol); bp 112-120°C/5mmHG. 7.7g (80%). <br><br> To a solution of 2.97g of pseudoionol in 10 ml of 5 acetonitrile, there were added, under stirring and while the temperature was kept below 30°C, 4.53g of triphenylphospine hydrochloride which had been obtained by passing dry hydrogen chloride into a solution of triphenylphosphine in dry ether. After the mixture had been left standing overnight at room temperature, 10 the acetonitrile was removed under reduced pressure below 50°C. To the residue there were added 4.47 gm of (+) S-6-Benzyloxy-2,5,7,8-tetramethylchroman-2-carbaldhyde in 15 ml of dimethylformamide, and the mixture was stirred. When a clear solution was obtained, sodium methoxide prepared from 0.352 g of 15 sodium and 7 ml of anhydrous methanol was stirred in, drop by drop below 15°C. The reaction mixture was turned red by the ylid formed. After the addition was complete, stirring was continued for 30 min at 10°C; then the mixture was gradually heated to 80°C, when the red color disappeared. The product was poured into 200 20 ml of 50% aqueous methanol, dried, and concentrated in vacuo. The residual oil was dissolved in 20 ml of ether, and an etheral solution of mercuric chloride was added until no more precipitate formed. When the precipitate was filtered and the filtrate was washed with water, dried and concentrated, to give 4.7 g of yellow 25 oil were obtained. The crude mixture of cis and trans alkene (MS (CI, m/z): 485.22 , M + H+, Calc. for C34H4402 484.7255) was dissolved in 30 ml of ethyl acetate and 0.80 g of 5% palladium on carbon was added, and the mixture was shaken under 40 psi of H2 for 30 hrs and then filtered through Celilte and rinsed well with <br><br> 3 6 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> ethyl acetate. The filtrate was concentrated and purified by silica gel chromatography eluting with EtOAc in hexane (1:9) to give 2,5,7,8-tetramethyl -(2R-(2RS,6RS,10-trimethylundecyl))-6- <br><br> chromanol (60% yield) XH-NMR (CDC13/TMS, ppm): 0.97 (m, 12H, 5 2a'-, 6a'-, 10a'-, ll'-CH3), 1.1 - 1.7 (m, 20H, 2'-, 6'-,10'-CH, l'-,3'-4'-,5'-,7,-,8'-,9,-CH2, 2a-CH3), 1.88 (m, 2H, 3-CH2), 2.17, 2.19, 2.20 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.63 (t, J = 6.7 Hz, 2H, 4-CH2); (MS (CI, m/z): 403.27 , M + H+, Calc. for C27H4602 402.6632. <br><br> A solution of 2,5,7,8-tetramethyl -(2R-(2RS,6RS,10-10 trimethylundecyl))-6-chromanol (0.466 g, 1.16 mmol) in N,N-dimethylformamide (20 mL) was treated with methyl bromoacetate (3.4 g, 8.3 mmol) and an excess of powdered NaOH (1.2 g, 3 0 mmol). The resulting yellow slurry was stirred vigorously for 24 h at room temperature. The reaction was acidified with 5 N HCl and 15 extracted with diethyl ether (3 x 30 ml). The combined ether layers were washed with H20 (3 x 30 ml) and brine (1 x 30 ml),and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel chromatography eluting with 19% (v/v) EtOAc and 2% acetic acid in hexanes. This yielded compound 20 17 in 76% yield. *H-NMR (CDC13/TMS, ppm): 0.97 (m, 12H, 2a'-, 6 a'-, 10a'-, ir-CH3), 1.2- 1.7 (m, 20H, 2'-, 6'-,10'-CH, l'-,3'- 4'-,5'-,7'-,8'-,9'-CH2, 2a-CH3), 1.92 (m, 2H, 3-CH2), 2.18, 2.20, 2.23 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.68 (t, J = 6.8 Hz, 2H, 4-CH2), 4.48 (s, 2H, OCH2); MS (CI, m/z): 461.44, M + H+, Calc. for C29H4804 460.700. <br><br> 25 <br><br> 37 <br><br> WO 01/58889 <br><br> PCT/USO 1/04168 <br><br> 2T5J7T8-t.etramftthyl-2R-(2,6T10-trimethyl-1 T3.5T9 deca tetraf»n^chroman-6-ylox v ) acetic acid (IS) <br><br> J2Z <br><br> 5 To a solution of methyl (-)-(S)- 6-hydroxy-2,5,7,8- <br><br> tetramethylchroman-2-carboxylate (20 gms 0.075mole) in 50ml of dry DMF, imidazole (13 gm, 0.191 lmole), and tert-butyldimethyl-silylchloride (14 gm, 0.0933 mole) were added. The mixture was stirred at 23°C for 24 hr and then treated with ether and poured 10 into IN HQ. The organic extracts were dried (brine, Na2S04) and concentrated in vacuo. The crude product was purified by flash chromatography (9:1 hexane :ethyl acetate) to yield 6-(dimethyl (1,1-dimethylethyl) silyl) - 2,5,7,8-tetramethyl-chroman-2-carboxylate (TBS protected methyl ester). . !H-NMR (CDC13/TMS, 15 ppm) : 0.12(s, 6H). 1.102(s, 9H), 1.18 (s, 3H), 1.48 (s, 3H), 1.645 (s, 3H), 2.07(s, 3H), 2.2 (t, J = 6.5hz 2H), 2.48-2.7 (m, 2H) and 3.72(s,3H, OCH3) (MS (CI, m/z): 379.32, M + H+, Calc. for C21H3404 378.586. <br><br> A solution of 3.78 g(10mmol) of the above ether ester, 20 in 20 ml of toluene and 10ml of CH2C12 was stirred with cooling from dry ice/acetone bath while 12 ml (18 mmol) of 25% disobutylaluminum hydride in toluene (Texas Alkyls) was added dropwise, over 10 min. After stirring at ca. -70° for 30 min, the reaction mixture was cautiously decomposed (-70°) with 10 ml of 25 MeOH. Following the addition of 50 ml of water and 50 ml of IN aqueous H2S04 solution, the mixture was warmed to RT, and worked <br><br> 38 <br><br> WO 01/58889 PCT/US01/04168 <br><br> up with ether in the usual way giving 3.2g (90%) of crude aldehyde ((+)S-6-(dimethyl(l,l-dimethylethyl)silyl)-2,5,7,8-tetramethyl-chroman-2-carbaldhyde) as a viscous oil which was purified by silica gel chromatography eluting with 19% (v/v) EtOAc in hexane. <br><br> 5 Concentration of the solution followed by drying under vacuo for 4 8 h yielded TBDS aldehyde (78%) as a solid of mp 66-68°C. *H-NMR (CDC13/TMS, ppm) : 0.12(s, 6H). l.l(s, 9H), 1.38 (s, 3H), 1.64 (s, 3H), 2.12 (s, 3H), 2.16(s, 3H), 2.3-2.2 (m, 2H), 2.53 (m, 2H) and 9.82(d, J=1.4Hz, 1H); MS (CI, m/z): 349.40 M + H+, Calc. for 10 C20H32SiO3 348.560. <br><br> To a solution of 2.97 g of psedoionol in 10 ml of acetonitrile, there were added, under stirring and while the temperature was kept below 30°C, 4.53 g of triphenylphospine hydrochloride which had been obtained by passing dry hydrogen 15 chloride into a solution of triphenylphosphine in dry ether. After the mixture had then been left standing overnight at room temperature, the acetonitrile was removed under reduced pressure below 50°C. To the residue there were added 4.80 gm of ((+)S-6-(dimethyl(l,l-dimethylethyl)silyl)-2,5,7,8-tetramethylchroman-2-20 carbaldhyde) in 15 ml of dimethylformamide, and the mixture was stirred. When a clear solution was obtained, sodium methoxide prepared from 0.352 g of sodium and 7 ml of anhydrous methanol was stirred in, drop by drop below 15°C. The reaction mixture was turned red by the ylid formed. After the addition was complete, 25 stirring was continued for 30 min at 10°C; then the mixture was gradually heated to 80°C, when the red color disappeared. The product was poured into 200 ml of 50% aqueous methanol, dried, and concentrated in vacuo. The residual oil was dissolved in 20 ml of ether, and an etheral solution of mercuric chloride was added <br><br> 39 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> until no more precipitate formed. When the precipitate was filtered and the filtrate was washed with water, dried and concentrated, to give 4.7 g of yellow oil were obtained. The crude silyl ether mixture of cis and trans alkene was dissolved in THF and tetra-n-5 butylammoniumfluoride (0.031mole) was added. After being stirred at 23°C for 40 minutes, the mixture was poured into water and extracted into ether. The ether extract was dried concentrated and purified by silica gel chromatography eluting with EtOAc in hexane (1:9) to give 2,5,7,8-tetramethyl-2R- (2,6,10-trimethyl-10 1,3,5,9 E:Z decatetraen)- 6-chromanol (68% yield). *H-NMR (CDC13/TMS, ppm) : 1.28 (s, 3H, 2aCH3), 1.65(s, 3H), 1.70(s,6H) 1.72 (s,3H), 1.9(m, 6H), 2.18 (s,3H), 2.35 (S, 6H), 2.53 (t, J = 6.6Hz, 2H, 4CH2), 5.13 - 5.27 (m, 3H) and 6.44(m, 2H) ; MS (CI, m/z): 395.17 M + H+, Calc. for C27H3g02 394.60. 15 A solution of 2,5,7,8-tetramethyl-2R- (2,6,10-trimethyl- <br><br> 1,3,5,9 E:Z decatetraen)- 6-chromanol (0.457 g, 1.16 mmol) in N,N-dimethylformamide (20 mL) was treated with methyl bromoacetate (3.4 g, 8.3 mmol) and an excess of powdered NaOH (1.2 g, 3 0 mmol). The resulting yellow slurry was stirred vigorously for 24 h 20 at room temperature. The reaction was acidified with 5 N HCl and extracted with diethyl ether (3 x 30 ml). The combined ether layers were washed with H20 (3 x 30 ml) and brine (1 x 30 ml), and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel chromatography eluting with 19% 25 (v/v) EtOAc and 2% acetic acid in hexanes. The resulting liquid was dissolved in diethyl ether (30 ml), washed with H20 (3 x 20 mL) and brine (1 x 20 mL), and then dried with Na2S04. The resulting solution was concentrated and dried in vacuo for 48 h. This yielded compound 18 in 67% yield. *H-NMR (CDC1/TMS, ppm) : 1.24 (s, <br><br> 40 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> 3H, 2aGH3), 1.63(s, 3H), 1.72(s,6H) 1.74 (s,3H), 1.92(m, 6H), 2.18 (s,3H), 2.29 (S, 6H), 2.43 (t, J = 6.6Hz, 2H, 4CH2), 4.68 (s,2H, OCH2), 5.10 - 5.27 (m, 3H) and 6.34(m, 2H) ; MS (CI, m/z): 452.24 M - H+, Calc. for C27H3802452.63. <br><br> 5 <br><br> 3-(2,5,7,8-tetramethy1-(2R-(4R,8,1 2-trimethyltridecyl) chroman-6-yloxy)propyl-l -ammonium chloride (19) <br><br> 10 A solution of 3-bromopropylamine hydrobromide (1.0 g, <br><br> 4.6 mmol) in a 2:1 dioxane/H20 (45 mL) was cooled to 0 °C and treated with K2C03 (6.22 g, 45 mmol) and di-tert-butyl dicarbonate (1.5 g, 6.9 mmol). The reaction was stirred for 15 h while warming to room temperature. The dioxane was removed in vacuo and the 15 remaining aqueous mixture was acidified with 5 N HCl and extracted with ethyl acetate (5 x 25 mL). The combined organic layers were dried with MgS04 and yielded 3-bromo-N-(tert-butoxycarbonyl)propylamine as a colorless oil (0.93 g, 93 %). 'H-NMR (CDC13/TMS, ppm): 1.41 (s 9H, CH3), 2.02 (quintet, J = 6.4 Hz, 20 2H, CH2), 3.23 (m, 2H, NCH2), 3.41 (t, J = 6.6 Hz, CH2Br), 4.8 (broad, 1H, NH); 13C-NMR (CDC13, ppm): 28.3 (CH3), 30.7, 32.6, 38.9 (CH2), 79.3 (quaternary C), 155.9 (CO); MS (CI, m/z): 239, 241 (M + H+Caic. for C8H16BrN02 237.03644). <br><br> A solution of R,R,R-a-tocopherol (0.5 g, 1.16 mmol) in 25 N,N-dimethylformamide (15 mL) was treated with 3-bromo-N-(tert-butoxycarbonyl)propylamine (0.9 g, 3.8 mmol) and an excess of <br><br> 41 <br><br> WO 01/58889 PCT/USO1/04168 <br><br> powdered NaOH (0.32 g, 8 mmol). The resulting yellow slurry was stirred vigorously for 24 h at room temperature. The reaction was acidified with 5 N HCl and extracted with diethyl ether (3 x 30 ml). The combined ether layers were washed with H20 (3 x 30 ml) and 5 brine (1 x 30 ml), and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel chromatography eluting with EtOAc (10% v/v) in hexanes. This yielded desired ether as a colorless oil (0.45 g, 66%). 'H-NMR (CDCI3/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 -10 1.6 (m, 33H, 4'-, 8'-, 12'-CH, 1'-, 2'-,3'-,5'-,6'-,7'-,9'-,10'-,ll'-CH2, 2a-CH3), 1.81 (m, 2H, 3-CH2), 1.99 (quintet, J = 6.2 Hz, 2H, CH2), 2.07, 2.14, 2.16 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.59 (t, J = 6.6 Hz, 2H, 4-CH2), 3.43 (m, 2H. NCH2), 3.73 (t, J = 5.7 Hz, 2H, OCH2), 4.34 (s, 2H, OCH2); 13C-NMR (CDC13, ppm): 11.7, 12.0, 12.9 (5a-, 7a-, 8 a-15 CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.7 (2a-CH3), 24.4, 24.8 (CH2), 27.9 (CH), 31.2 (3-CH2), 32.7, 32.8 (CH), 37.2, 37.4, 37.5, 39.3, 40.1 (CH2), 70.2 (OCH2), 74.8 (2-C), 117.5, 122.9, 125.5, 127.5 (aryl C), 147.5, 148.0 (aryl C-O), 156.0 (CO); MS (CI, m/z): 589 M + H+, Calc. for C37H65N04 587.49136. 20 The above N-protected ether (0,1 g, 0.17 mmol) was dissolved 4 N HCl in dioxane (1 mL, 4 mmol) and stirred for 4 h. The dioxane was removed by blowing a stream of argon over the reaction mixture. The resulting material was dried in vacuo for 8 h yielding 19 as a white solid (82 mg, 99%). VH-NMR (CDC13/TMS, 25 ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 33H, 4'-, 8'-, 12'-CH, 1'-, 2'-, S'-^'-^'-J'-^'-.lO'-Jl'-CH,, 2a-CH3), 1.81 (m, 2H, 3-CH2), 1.99 (quintet, J = 6.2 Hz, 2H, CH2), 2.07, 2.11, 2.15 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.29 (m, 2H, CH2), 2.59 (t, J = 6.6 Hz, 2H, 4-CH2), 3.43 (m, 2H. NCH2), 3.79 (m, 2H, OCH2) I3C-NMR <br><br> 42 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> (CDCI3, ppm): 11.8, 11.9, 12.7 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.9 (2a-CH3), 24.4, 24.8 (CH2), 28.0 (CH), 28.4 (CH3), 31.2 (3-CH2), 32.7, 32.8 (CH), 37.3, 37.4, 37.5, 39.4, 40.0 (CH2), 74.8 (OCH2), 75.0 (2-C), 117.5, 122.9, 126.0, 127.3 (aryl C), 147.8, 148.0 (aryl C-O); HRMS (CI, m/z): 487.438887 (M + H+, Calc. for C32H57N02 487.438935). <br><br> 10 <br><br> 2,5,7,8 - tetr a m eth y 1 - (2R - (4R T 8 R, 12 - tri m eth y 1 tri d ecy 1) chroman-3-ftnft-6-y1oyy) acetic acid (20) <br><br> A solution of R,R,R,-a-tocopherol acetate (2 g, 4.2mmol) in anhydrous toluene (150 mL) was heated to reflux and then treated with 2,3-dichloro-5,6-dicyano-l,4-benzoquinone (0.96 g, 15 4.2 mmol) in 4 portions at 1 h intervals. The reaction was refluxed for 24 h. During this time the reaction mixture became a dark red color and then it precipitated a light colored solid. The reaction was cooled to room temperature, filtered, and the filtrate was concentrated. The resulting dark colored oil was purified by silica 20 gel chromatography eluting with ethyl acetate (10%, v/v) in hexanes. This yielded the desired chromene acetate as a colorless oil (1.74g, 88%). JH-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-, 12'-CH, l'-,2'-,3'-,5'-,6'-,7'-,9'-,10'-,ll'-CH2, 2a-CH3), 2.07, 2.13, 2.18 (3 x s, 9H, 5a-, 25 7a-, 8a-CH3), 2.35 (s, 3H, CH3CO-), 5.61, 6.52 (2 x d, J = 10:0 Hz, 2H, CH); 13C-NMR (CDC13, ppm): 11.5, 11.6, 13.1 (5a-, 7a-, 8a-CH3), <br><br> 43 <br><br> WO 01/58889 PCT/US01/04168 <br><br> 14.1 (CH3), 19.6, 19.7 (CH3), 20.4, 21.4 (CH2), 22.6, 22.7 (CH3), 24.4, 24.8 (CH2), 25.8 (2a-CH3), 27.9 (CH), 30.8 (3-CH2), 32.7, 32.8 (CH), 37.2, 37.4, 39.4, 41.0 (CH2), 60.3 (2-C), 117.6, 119.7, 122.3, 122.6, 128.9, 129.6 (aryl and vinyl C), 141.2, 148.4 (aryl C-O), 5 169.4 (CO); HRMS (CI, m/z): 471.375799 M + H+, Calc. for C3iH5o03 470.375996. <br><br> A solution of the chromene acetate (1.0 g, 2.13 mmol) in ethanol (20 mL) was treated with 2 N NaOH (20 mL) and stirred at 60 °C for 90 min. The reaction mixture was cooled, acidified 10 with 5 N HCl, and the ethanol was removed in vacuo. The resulting aqueous solution was extracted with ether and concentrated to a light yellow oil that was purified by silica gel chromatography eluting with ethyl acetate (15%, v/v) in hexanes. This yielded the desired chromene-6-ol intermediate as a colorless oil (0.92 g, 98%). 15 'H-NMR (CDC1/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13!-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-, 12'-CH, l,-,2,-,3,-,5,-,6'-, 7,-,9'-,10'-,H'-CH2, 2a-CH3), 2.14, 2.18, 2.19 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 5.63, 6.55 (2 x d, J = 10.0 Hz, 2H, CH); 13C-NMR (CDC13, ppm): 10.8, 11.6, 12.4 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 21.3 (CH2), 22.6, 20 22.7 (CH3), 24.4, 24.8 (CH2), 25.2 (2a-CH3), 27.9 (CH), 30.9 (3-CH2), 32.7, 32.8 (CH), 37.2, 37.4, 37.5, 39.3, 40.5 (CH2), 50.8 (2-C), 116.2, 117.8, 120.1, 122.3, 123.0, 130.0 (aryl and vinyl C), 144.6, 145.3 (aryl C-O), 169.4 (CO); HRMS (CI, m/z): 428.365275 M + H-, Calc. for C29H4802 428.365431. <br><br> 25 A solution of the chromene-6-ol intermediate (0.9 g, 2.1 <br><br> mmol) in N,N-dimethylformamide (20 mL) was treated with methyl bromoacetate (3.4 g, 8.3 mmol) and an excess of powdered NaOH (1.2 g, 30 mmol). The resulting yellow slurry was stirred vigorously for 24 h at room temperature. The reaction was acidified with 5 N <br><br> 44 <br><br> WO 01/58889 PCT/US01/04168 <br><br> HCl and extracted with diethyl ether (3 x 30 ml). The combined ether layers were washed with H20 (3 x 30 ml) and brine (1x30 ml), and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel 5 chromatography eluting with 19% (v/v) EtOAc and 2% acetic acid in hexanes. The resulting yellow liquid was dissolved in diethyl ether (30 ml), washed with H20 (3 x 20 mL) and brine (1 x 20 mL), and then dried with Na2S04. The resulting solution was concentrated to a light yellow oil and dried in vacuo for 48 h. This yielded 19 as a 10 colorless (0.90 g, 88%). ^-NMR (CDCI3/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a1-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-, 12'-CH, 1'-, 2'-,3'-,5'-,6'-, 7'-,9'-,10,-,ir-CH2, 2a-CH3), 2.07, 2.10, 2.19 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 4.37 (s, 2H, OCH2), 5.62, 6.50 (2 x d, J = 10.0 Hz, 2H, CH); 13C-NMR (CDC13, ppm): 11.3, 11.5, 12.9 (5a-, 7a-, 8a-CH3), 15 19.6, 19.7 (CH3), 21.3 (CH2), 22.6, 22.7 (CH3), 24.4, 24.8 (CH2), 25.6 (2a-CH3), 27.9 (CH), 30.9 (3-CH2), 32.7, 32.8 (CH), 37.2, 37.4, 37.5, 39.3, 40.9 &lt;CH2), 60.5 (OCH2), 69.1 (2-C), 118.0, 119.8, 122.8, 122.9, 129.6, 19.8 (aryl and vinyl C), 147.5, 147.8 (aryl C-O), 173.4 (CO); HRMS (CI, m/z): 487.378731 M + H+, Calc. for 20 C31HSi04 487.378736. <br><br> 2-(2,5,7,8-tetramethy1-(2R-(4R,8,12-tnmethy1tridecy1) chroman-6-y1oxy)tri ethyl ammonium sulfate (21) <br><br> Et3NH 03S0&gt;s^/\ <br><br> 25 <br><br> 45 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> A solution of 2-(2,5,7,8-tetramethyl-(2R-(4R,8R,12-trimethyltridecyl)chroman-6-yloxy))ethan-l-ol (13) (0.1 g, 0.21 mmol) in anhydrous DMF (2 mL) and pyridine (0.6 mL) was treated sulfur trioxide-N,N-dimethylformamide complex (0.16 g, 1.0 mmol), 5 and the resulting solution was stirred for 24 h. The reaction mixture was quenched with 1 N HCl and then extracted with CH2C12 (5x5 mL). Gaseous ammonia was bubbled through the CH2C12 solution for 10 min. The resulting solution was concentrated to a yellow paste and purified by silica gel chromatography eluting with 10 MeOH (10%, v/v) and triethyl amine (2%) in CHC13. This yielded 21 as a yellow semi-solid (92 mg, 77%) TC-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 33H, 4'-, 8'-,12'-CH, l'-^'-^'-^'-^'-J'-^'-.lO'-Jl'-CH,, 2a-CH3), 1.81 (m, 2H, 3-CH2), 1.95 2.01, 2.05 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.45 (t, J = 6.6 15 Hz, 2H, 4-CH2), 3.05 (m, 6H, NCH2), 3.79 (m, 2H, OCH2), 4.21 (m, 2H, OCH2); 13C-NMR (CDC13, ppm): 9.46 (CH3), 12.4, 12.6, 13.5 (5a-, 7a-, 8a-CH3), 20.3, 20.4 (CH3), 21.3, 21.7 (CH2), 23.3, 23.4 (CH3), 24.5 (2a-CH3), 25.1, 25.5 (CH2), 28.6 (CH), 31.9 (3-CH2), 33.3, 33.4 (CH), 37.9, 38.1, 40.0, 40.8 (CH2), 46.9 (NCH2), 67.4, 71.9 (OCH2), 20 75.5 (2-C), 118.3, 123.5, 126.5, 128.3 (aryl C), 148.5 (aryl C-O); HRMS (CI, m/z): 554.364102 M - NH3 , Calc. for C31H5406S 554.364119. <br><br> 6-C2,5,7,8-tfttramftthy1-(7.R-(4R ,8,1 2-t.rim ethyl tri rie.cyl) 25 f.hrnman) aceti c acid (2.2..) <br><br> 46 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> A solution of R,R,R-a-tocopherol (1.0 g, 2.3 mmol) in anhydrous CH2C12 (25 mL) was cooled to O °C. Diisopropylethyl amine (2 mL, 11.6 mmol) was added followed by the drop wise addition of trifluoromethylsulfonic anhydride (5.0 g, 17.7 mmol). <br><br> 5 The solution turned to a dark immediately and was allowed to warm to room temperature while stirring for 24 h. The reaction was quenched with H20 and then was extracted with diethyl ether (2 x 100 mL). The combined ether layers were washed with 1 N HCl (50 mL), H20 (50 mL), brine (50 mL), and then dried with MgS04. The 10 ether solution was concentrated to a yellow oil and purified by silica gel chromatography eluting with ethyl acetate (3%, v/v) in hexane. This yielded the desired triflate intermediate as a yellow oil (1.3 g, quantitative). ^-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, 4a*-, 8a*-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-, 12'-CH, 1'-, 2'-,3'-,5*-,6'-15 ,7,-,9,-,10'-,ll'-CH2, 2a-CH3), 1.81 (m, 2H, 3-CH2), 2.07, 2.13, 2.21 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.62 (t, J = 6.6 Hz, 2H, 4-CH2); 13C-NMR (CDC13, ppm): 11.9, 13.2, 14.0 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.8 (CH2), 28.0 (CH), 31.2 (3-CH2), 32.7, 32.8 (CH), 37.3, 37.4, 37.5, 20 39.4, 40.0 (CH2), 75.6 (2-C), 118.4, 124.4, 126.7, 128.1 (aryl C), 139.6, 150.9 (aryl C-O); 19F~NMR (CDC13, ppm): -73.52; HRMS (CI, m/z): 563.337803 (M + H+, Calc. for C3oH5o04F3S 563.338192). <br><br> A solution of the triflate (1.3 g, 2.31 mmol) in anhydrous DMF (23 mL) was treated with LiCl (0.98 g, 4.62 mmol), 25 triphenylphosphine (0.37 g, 1.4 mmol), 2,6-di-tert-butyl-4-methylphenol (2-3 crystals), tributyl(vinyl)tin (0.73 g, 2.31 mmol), and dichlorobis(triphenylphosphine)-palladium(II) (0.24 g, 0.35 mmol). This mixture was heated to 120 °C and stirred. After 2h, additional tributyl(vinyl)tin (0.73 g, 2.31 mmol). After 8 h, the <br><br> 47 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> reaction was cooled to room temperature and added to a mixture of H20 (50 mL) and diethyl ether (50 mL). The ether layer was washed with 1 N HCl (6 x 30 mL) and a saturated solution of KF (6 x 30 mL). The ether solution was dried with Na2S04 and then concentrated to a 5 dark oil. This material was purified by silica gel chromatography eluting with ethyl acetate (3%, v/v) in hexane yielding the 6-vinylchroman intermediate as a clear oil (0.38 g, 38%). 'H-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a*-, 12a'-, 13'-CH3), 1.0 - <br><br> I.6 (m, 24H, 4'-, 8'-, 12'-CH, <br><br> 10 2a-CH3), 1.86 (m, 2H, 3-CH2), 2.20, 2.24, 2.28 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.62 (t, J = 6.8 Hz, 2H, 4-CH2), 5.18, 5.56 (2 x dd, Jgem = 2.3 Hz, Jcis = 11.2 Hz, Jtran8 = 18.7 Hz, 2H, =CH2), 6.77 (dd, J = 18.7, <br><br> II.2 Hz, 1H, CH); I3C-NMR (CDC13, ppm): 11.9, 16.3, 17.2 (5a-, 7a-, 8a-CH3), 19.7, 19.8 (CH3), 20.8, 21.1 (CH2), 22.6, 22.7 (CH3), 23.9 <br><br> 15 (2a-CH3), 24,5, 24.8 (CH2), 28.0 (CH), 31.2 (3-CH2), 32.7, 32.8 (CH), 37.3, 37.5, 37.5, 39.4, 40.1 (CH2), 74.9 (2-C), 116.7, 119.0, 122.0, 129.8, 131.2, 132.8, 136.8 (aryl/vinyl C), 150.9 (aryl C-O); HRMS (CI, m/z): 440.401602 (M + H+, Calc. for C31H520 440.401812). <br><br> 20 A solution of the 6-vinylchroman intermediate (0.12 g, <br><br> 0.27 mmol) in anhydrous THF (1 mL) was cooled to 0 °C and treated with 9-BBN (0.60 mL, 0.5 M in THF, 0.3 mmol). The reaction mixture was heated to reflux for 8 h. The reaction was quenched with water (1.5 mL) and treated with NaB03*4H20 and the resulting 25 slurry was stirred overnight. Diethyl ether (4 mL) and the reaction mixture were extracted with CH2C12 (2 x 20 mL). The organic layers were concentrated to a clear oil that was purified by silica gel chromatography eluting with ethyl acetate (50%, v/v) in hexane. This yielded the desired 6-(2-hydroxyethyl)chroman intermediate as <br><br> 48 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> a colorless oil (30 mg, 24 %). !H-NMR (CDC13/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8'-, 12'-CH, l'-^'-^'-^'-^'-J'-^'-aO'-.H'-CHz, 2a-CH3), 1.81 (m, 2H, 3-CH2), 2.17, 2.24, 2.28 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.68 (t, J = 6.8 Hz, 2H, 5 4-CH2), 3.01 (t, J = 7.5 Hz, 2H, Ar-CH2), 3.74 (t, J = 7.5 Hz, 2H, OCH2); 13C-NMR (CDC13, ppm): 12.0, 15.1, 16.0 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.8 (CH2), 28.0 (CH), 31.2 (3-CH2), 32.7, 32.8 (CH), 37.3, 37.4, 37.5, 39.4, 40.0 (CH2), 62.2 (OCH2), 72.6 (2-C), 116.8, 122.3, 10 124.9, 132.4, 133.9 (aryl C), 150.1 (aryl C-O); HRMS (CI, m/z): 458.412154 (M + H+, Calc. for C31H5402 458.412384). <br><br> A solution of pyridinium chlorochromate (32 mg, 0.1 mmol) in anhydrous CH2C12 (0.5 mL) was treated with a solution of the 6-(2-hydroxyethyl)chroman intermediate (32 mg, 0.07 mmol) in 15 CH2C12 (0.5 mL). The reaction was stirred for 2 h at which time n o starting material was visible by thin layer chromatography. Diethyl ether (2 mL) was added and the resulting solution was filtered through a thin pad of celite. The filtrate as concentrated and yielded a yellow oil (20 mg). This oil was dissolved in t-BuOH (0.5 20 mL) and treated with phosphate buffer (0.5 mL, 1 N, pH = 4.0), 2-methyl-2-butene (0.1 mL) and NaC102 (5.4 mg, 0.05 mmol). After stirring for 40 min, the reaction mixture was extracted withCHCl3 (6 x 10 mL) and the combined organic layers were dried with Na2S04. The CHC13 solution was concentrated to a yellow oil that was 25 purified by preparative thin layer chromatography eluting with ethyl acetate (30%, v/v) and acetic acid (1%) in hexanes. This yielded 2 2 as colorless oil (20 mg, 63%). 'H-NMR (CDC1/TMS, ppm): 0.87 (m, 12H, 4a'-, 8a'-, 12a'-, 13'-CH3), 1.0 - 1.6 (m, 24H, 4'-, 8', 12'-CH, 1'-^'-^'-.S'-^'-J'-^XlOXll'-CH,, 2a-CH3), 1.81 (m, 2H, 3-CH2), <br><br> 49 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> 2.17, 2.24, 2.28 (3 x s, 9H, 5a-, 7a-, 8a-CH3), 2.66 (t, J = 6.8 Hz, 2H, 4-CH2), 3.71 (s, 2H, CH2COOH); I3C-NMR (CDC13, ppm): 12.0, 15.3, 16.2 (5a-, 7a-, 8a-CH3), 19.6, 19.7 (CH3), 20.6, 21.0 (CH2), 22.6, 22.7 (CH3), 23.8 (2a-CH3), 24.4, 24.8 (CH2), 28.0 (CH), 28.9, 31.2 5 (3-CH2), 32.7, 32.8 (CH), 37.3, 37.4, 37.5, 39.4, 40.0 (CH2), 72.6 (2-C), 117.1, 122.2, 124.9, 132.4, 132.7 (aryl C), 150.2 (aryl C-O), 179.2 (COOH); HRMS (CI, m/z): 472.391583 (M + H+, Calc. for C31H5203 472.391644). <br><br> 10 2 T 5,7,8 - t.p.tr a m eth y 1 - f 2K - (h ep t.v n chroman-6-yloxy)acetic acid (23) <br><br> V <br><br> o <br><br> A solution of hexyltriphenyphosphonium bromide (0.880g, 2.05mmol) in 11.2 ml of anhydrous DME was stirred at 15 room temperature while 0.86 ml (2.06mmol) of 2.4 M n-butyllithium in hexane was added. The resulting red solution was stirred for 2h at room temperature, then a solution of ((+)S-6-Benzyloxy-2,5,7,8-tetramethylchroman-2-carbaldhyde (306 mg, 0.944 mmol) in 3 ml of anhydrous DME was added and stirring was 20 continued for 3 h at 65-75°C. After cooling, the reaction mixture was poured into cold dilute H2S04 and work up ether was carried out in the usual manner. The ether was concentrated in vacuo to afford the oily material. Product was isolated using column chromatography and eluted with chloroform to yield 46% of the 25 product. The mixture of cis and trans alkene was dissolved in 30 ml of ethyl acetate and 50 mg of 5% palladium on carbon was added, <br><br> 50 <br><br> WO 01/58889 <br><br> PCT/USO 1/04168 <br><br> and the mixture was shaken under 40 psi of H2 for 10 hrs and then filtered through Celilte and rinsed well with ethyl acetate. The filtrate was concentrated and purified by silica gel chromatography eluting with EtOAc in hexane (1:9) to give (2R) 2,5,7,8-tetramethyl-5 2-(heptyl)-6-chromanol (60% yield) *H-NMR (CDC13/TMS, ppm):0.89 (s, 3H), 1.3-1.5 (m, 15H), 1.89 (m, 2H), 2.2 (s, 3H), 2.08(s, 3H), 2.23 (s, 3H) , and 2.48 (t, J =6.5 Hz, 2H); MS (CI, m/z):305.35 M + H+, Calc. for C2oH3202304.4746). <br><br> A solution of 2,5,7,8-tetramethyl-2-(heptyl) chromanol 10 (0.353 g, 1.16 mmol) in N,N-dimethylformamide (20 mL) was treated with methyl bromoacetate (3.4 g, 8.3 mmol) and an excess of powdered NaOH (1.2 g, 30 mmol). The resulting yellow slurry was stirred vigorously for 24 h at room temperature. The reaction was acidified with 5 N HCl and extracted with diethyl ether (3x30 15 ml). The combined ether layers were washed with H20 (3 x 30 ml) and brine (1 x 30 ml), and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by silica gel chromatography eluting with 19% (v/v) EtOAc and 2% acetic acid in hexanes. The resulting liquid was dissolved in diethyl ether 20 (30 ml), washed with H20 (3 x 20 mL) and brine (1 x 20 mL), and then dried with Na2S04. The resulting solution was concentrated and dried in vacuo for 48 h. This yielded compound 23 in 36%yield. !H-NMR (CDC1/TMS, ppm): 'H-NMR (CDC13/TMS, ppm): 0.88 (s, 3H), 1.2-1.5 (m, 15H), 1.88 (m, 2H), 2.1 (s, 3H), 2.18(s, 25 3H), 2.2 (s, 3H) , 2.55 (t, J =6.5 Hz, 2H) and 4.78 (s, 2H); HRMS (CI, m/z):363.2535 (M + H+, Calc. for C22H3504363.2541). <br><br> 51 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> 2.5.7. R -te.tr a m pt.hy 1 -f2R -(tri d p.cy 1) chroman-6-y1r&gt;xy^ <br><br> acetic acid (24) <br><br> The compounds 24 and 25 were synthesized in manner identical to the synthesis of 23 using appropriate phosphonium 5 bromide. <br><br> !H-NMR (CDCI3/TMS, ppm): 0.83 (s, 3H), 1.25-1.57 (m, 10 27H), 1.88 (m, 2H), 2.1 (s, 3H), 2.18 (s, 3H), 2.20(s, 3H), 2.55 (t, J =6.6 Hz, 2H) and 4.48 (s, 2H) ; MS (CI, m/z): 447.14 M + H+, Calc. For C28H4604 446.6732. <br><br> 9,5,7. R-tp.tr am p.thvl -(?.R - (h p.pta d ecy l) chroman-6-y1oxy)acet.ic anid <br><br> 15 (25) <br><br> 'H-NMR (CDCI3/TMS, ppm): 0.86 (s, 3H), 1.15-1.67 (m, 20 35H), 1.88 (m, 2H), 2.16 (s, 3H), 2.20 (s, 3H), 2.23(s, 3H), 2.55 (t, J =6.4 Hz, 2H) and 4.78 (s, 2H) ; MS (CI, m/z): 503.45 M + H+, Calc. For C32H54O4 502.781. <br><br> 52 <br><br> WO 01/58889 PCT/US01/04168 <br><br> ?. T 5 r 71R - tp.tr a m eth y 1-f 4 T R, -rl i m p.th y 1 -1.3 T 7 EZ <br><br> nnnntrien) chromnn-6-y1oxy^ acetic arid <br><br> Compouna 26 was synthesized in a manner identical to the synthesis of compound 18 using nerol instead of pseudoionol. <br><br> 10 XH-NMR (CDCI3/TMS, ppm) : 1.24 (s, 3H, 2aCH3), 1.63(m, 1H), 1.68 (s,3H), 1.74(s,6H), 1.92(m, 6H), 2.18 (s,3H), 2.29 (S, 6H), 2.43 (t, J = 6.6Hz, 2H, 4CH2), 4.68 (s, 2H, OCH2), 5.64(m, 2H) and 5.27 (m, 1H) ; MS (CI, m/z): 413.24 M +H+, Calc. for C26H3604412.0115. <br><br> 15 R.Z, RS, RS, RS-(phyty1trimethy1hen7.enethiol-6-y1oxy) <br><br> 2,3,6-trimethylphenol (1.6 g, 11.8 mmol). was dissolved in 50 mL of anhydrous methanol which had been deoxygenated by bubbling with nitrogen. Ammonium thiocyanate (2.2 g, 28,9 mmol) 25 was added to this solution which was then cooled to 0 °C and bubbled with chlorine gas. The initially colorless homogeneous solution becomes pink and then green with the formation of a white precipitate. The solution was stirred for 1 h at 0 0 C and then for a further hour at 20 0 C. The dissolved chlorine was removed by <br><br> 53 <br><br> WO 01/58889 <br><br> PCT/USO 1/04168 <br><br> bubbling with nitrogen and the precipitate removed by filtration. Evaporation of the filtrate under reduced pressure followed by drying under high vacuum (0.1 torr) yielded 2.20 g (97%) of 2,3,5-Trimethyl-4-hydroxyphenylthiocyanate in a form pure enough for 5 the next step in the synthesis. An analytical sample was recrystallized from hexanes : white crystals, mp 100.3 0 C. *H NMR (CDC13) 8 7.2 (s, 1 H), 5.0 (s, 1 H) 2.4 (s, 3 H), 2.2 (s, 6 H). <br><br> 2,3,5-Trimethyl-4-hydroxyphenylthiocyanate (2 g, 10.35 mmol) was dissolved in 100 mL of anhydrous ether containing 25 10 mL of anhydrous tetrahydrofuran. This solution was added dropwise over 1 h to 100 mL of anhydrous ether containing LiAlH 4 (0.9 g, 24 mmol) at room temperature. After a further hour at 20 0 C, the unreacted LiAlHi was destroyed by cooling the heterogeneous mixture to 0 °C and adding moist ether (50 mL), H20 (50 mL), and 1 15 N HCl (50 mL). A further 50 mL of water was added and the organic phase was separated and washed with water (2 x 50 mL), NaHC03 solution (2 x 50 mL), water (2 x 50 mL), and saturated NaCl (50 mL). The organic phase was dried over anhydrous MgS04 and filtered and the solvent removed under reduced pressure. Silica gel 20 column chromatography with 5% ethyl acetate in hexane gave 1.8 g (90%) of 2,3,5-trimethyl-4-hydroxybenzenethiol as a white powder, mp 86 °C (Lit.1 mp 86 °C). <br><br> Solution of 2,3,5-trimethyl-4-hydroxybenzenethiol (3 g, 17.83 mmol ), isophytol (4.8 g, 16.19 mmol), anhydrous zinc 25 chloride (1.2 g, 8.8 mmol) and 0.2 mL of glacial acetic acid in 3 0 mL of absolute ether was refluxed for 1 h. The solvent was then removed in vacuo at 50° C and the red oil obtained was dissolved in a mixture of 50 mL of petroleun ether and 20 mL of 70% aqueous methanol. The ether layer was dried (Na2S04) and evaporated in <br><br> 54 <br><br> WO 01/58889 PCT/USO1/04168 <br><br> vacuo to give a red oil, which was purified by silica gel chromatography eluting with hexans:ether (9:1) to give 3g(38%) E.Z, RS, RS, RS-Phytyltrimethylhydroxybenzenethiol as yellow oil. JH NMR (CDC13) 87.11 (s, 1 H, Ar-H), 5.23 (t, 1 H, vinylic-H), 4.62 (s, 1 5 H, OH), 3.34 (d, 2 H, Ar-S-CH2-), 2.41 (s, 3 H, Ar-CH3), 2.19 (s, 3 H, Ar-CH3), 2.18 (s, 3 H, Ar-CH3), 0.83-1.92 (m, 39 H, Phytol chain). <br><br> A solution of phytyltrimethylhydroxybenzenethiol (3g, 6.7 mmol) in N, N-dimethyl-formamide (80 mL) was treated with methyl bromoacetate (7.4 g, 48.3 mmol) and an excess of powdered 10 NaOH (7 g, 175 mmol). The resulting pink oil was stirred at RT for 24 h. The reaction mixture was acidified with 5 N HCl and extracted with ether (3 x 150 mL). The combined ether layers were washed with H20 (3 x 150 mL) and brine (1 x 150 mL), and then dried (Na2S04). The ether solution was concentrated to a yellow oil that 15 was purified by silica gel chromatography eluting with 20% EtOAc in hexane to give 3 g (88%) of E.Z, RS, RS, RS-(phytyltrimethylbenzenethiol-6-yloxy)acetic acid as a yellow oil. lH NMR (CDC13) 5 10.90 (s, 1 H, COOH), 8.08 (s, 1 H, Ar-H), 5.30 (t, 1 <br><br> H, vinylic-H), 4.35 (s, 2 H, CH2COOH), 3.42 (d, 2 H, Ar-S-CH2-), 2.34 20 (s, 3 H, Ar-CH3), 2.25 (s, 3 H, Ar-CH3), 2.22 (s, 3 H, Ar-CH3), 0.83- <br><br> I.94 (m, 39 H, Phytyl chain). HRMS (CI, m/z): 504.362821 ( M+H\ Calc. for C31H5303S 504.363718). <br><br> (R)-2((2,5,7,8-tetramethyl-2-(3 propene methyl ester) <br><br> 25 chroman-6-vIoxy)acetic acid (28.) <br><br> 55 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> To a slurry of (carbomethoxymethyl)triphenyl phosphonium bromide (1.8 gm, 4.32 mmol) in 12 ml of THF at °C was added 1.66 ml of n-BuLi (2.5M in hexane) dropwise. The resulting solution was removed to room temperature for 2h, and 5 then a solution of (+)S-6-(dimethyl(l,l-dimethylethyl)silyl)-2,5,7,8-tetramethyl chroman-2-carbaldhyde (1.31g, 3.76 mmol) in 7 ml THF was added via cannula. The solution was stirred at room temperature for 44hr and then 10 ml of IN aq. HCl was added. The layer were separated and then aq. phase was extracted with ether ( 10 3 X 15 ml). The combined organic layer were washed with brine, dried over Na2S04 and filtered. After concentration of the filtrate, the crude alkene was purified by flash chromatography eluting with dichloromethane to give mixture of the cis and trans alkene in 93% yield. The silyl ether mixture of cis and trans alkene (3.76mmol) 15 was dissolved in THF and tetra-n-butylammoniumfluoride (0.041mole) was added. After being stirred at 23°C for 1.5h, the mixture was poured into water and extracted into ether, The ether extract was dried concentrated and purified by silica gel chromatography eluting with EtOAc in hexane (3:7) and both the cis 20 and trans isomer of 2,5,7,8-tetramethyl-2R-(3'propenemethyl ester)-6-chromanol were isolated and characterized (68% yield) *H-NMR (CDC1/TMS, ppm): 1.65 (s, 3H, 2a CH3), 2.12 (m, 2H, 3CH2), 2.39 (s, 9H, CH3), 2.48 (m, 2H, 4 CH2), 3.78 (s, 3H, OCH3), 6.11 (d, 1H, CH=) and 7.13 (d, 1H, CH=). <br><br> 25 A solution of 2,5,7,8-tetramethyl-2R-(3'propene methyl ester)6-chromanol (0.353 g, 1.16 mmol) in N,N-dimethylformamide (20 mL) was treated with methyl bromoacetate (3.4 g, 8.3 mmol) and an excess of powdered NaOH (1.2 g, 30 mmol). The resulting yellow slurry was stirred vigorously for 24 h at room temperature. <br><br> 56 <br><br> WO 01/58889 <br><br> PCT/USO1/04168 <br><br> The reaction was acidified with 5 N HCl and extracted with diethyl ether (3 x 30 ml). The combined ether layers were washed with H20 (3 x 30 ml) and brine (1 x 30 ml), and then dried with Na2S04. The ether solution was concentrated to a yellow oil that was purified by 5 silica gel chromatography eluting with 19% (v/v) EtOAc and 2% acetic acid in hexanes. The resulting liquid was dissolved in diethyl ether (30 ml), washed with H20 (3 x 20 mL) and brine (1 x 20 mL), and then dried with Na2S04. The resulting solution was concentrated and dried in vacuo for 48 h. This yielded compound 10 28 in 40%yield. JH-NMR (CDC13/TMS, ppm): 1.68 (s, 3H, 2a CH3), 2.11 (m, 2H, 3CH2), 2.36 (s, 9H, CH3), 2.56 (m, 2H, 4 CH2), 3.70 (s, 3H, OCH3), 4.78 (s, 2H, OCH2), 6.03 (d,lH, CH=) and 7.03 (d,lH, CH=); MS (CI, m/z):337.24 M+H+, Calc. for ClgH2406 336.3867 . <br><br> 15 2.5T7,8-tetramethy1-(2R-(propionate) chroman-6-yloxy) <br><br> acetic ar.iri (29) <br><br> 20 <br><br> B <br><br> The mixture of cis and trans alkene 2,5,7,8-tetramethyl-2R-(3 propene methylester)-6-chromanol was dissolved in 30 ml of ethyl acetate and 50 mg of 5% palladium on carbon was added, and 25 the mixture was shaken under 40 psi of H2 for 24 hrs and then filtered through Celilte and rinsed well with ethyl acetate. The filtrate was concentrated and purified by silica gel chromatography eluting with EtOAc in hexane (1:9) to give compound # 29. XH-NMR (CDCI3/TMS, ppm): 1.62 (s, 3H, 2a CH3), 2.0-2.3 (m, 6H, CH2), 2.41 <br><br> 57 <br><br> 10 <br><br> WO 01/58889 PCT/USO1/04168 <br><br> (s, 9H, CH3), 2.53 (m, 2H, 4 CH2), 3.67 (s, 3H, OCH3) and 4.88 (s, 2H, OCH2); MS (CI, m/z):339.34 M+H+, Calc. for C18H26Q6338.4025. <br><br> EX A MPT ,F. 3 <br><br> Synthesis of all-racftmic 1-aza-nr-tonophftrol analogs: <br><br> PhytyHrimftthylhydrogninnnft (31) <br><br> C^5H33 <br><br> The solution of trimethylhydroquinone (30, 7.5 g, 49.28 15 mmol), isophytol (12 g, 40.47 mmol), anhydrous zinc chloride (3 g, 22.01 mmol) and 0.4 mL of glacial acetic acid in 60 mL of absolute ether was refluxed for 1 h. The solvent was then removed in vacuo at 50° C and the slurry obtained was dissolved in a mixture of 50 mL of petroleum ether and 20 mL of 70% aqueous methanol. The 20 emulsion formed was destroyed by the addition of 20 mL of ether. The ether layer was dried (Na2S04) and evaporated in vacuo to give a red solid paste that was triturated vigorously with 70 mL of petroleum ether. After cooling to -78°C, the suspension was centrifuged and the supernatant petroleum ether was decanted. 25 Cold petroleum ether (10 mL) was added to the crystalline mass, and the operation was twice repeated to give 10 g (47%) of sticky white solid, which was used in the next step without further purification. <br><br> 30 <br><br> 58 <br><br> WO 01/58889 PCT/US01/04168 <br><br> Phytyltrimp.thylhyrirngiiinonp. diacetate—(32) <br><br> CiftH <br><br> 16n33 <br><br> OAc <br><br> The crude phytyltrimethylhydroquinone (31, 9 g, 20.1 mmol) was dissolved in dry pyridine (90 g) and acetic anhydride (90 g). After storage for 10 h at room temperature, the mixture 10 was poured onto ice (200 mL) and extracted with ether (2 x lOOmL). The ether solution was washed with 3N sulfuric acid (2 x 100 mL), 10% sodium bicarbonate solution (2 x lOOmL), and again water (2 x 100 mL), dried (Na2S04), and evaporated to yield 10.2g (95%) of diacetate (32) as a light yellow oil. The oil was used in the 15 next step without further purification. <br><br> Trimp.fhy1(3.7-1 1.1 5-tetramet.hy1-3-henzamiHnhexariftpyl) hydroquinone diacetate (33) <br><br> 20 <br><br> 25 <br><br> I OAc NH <br><br> cieHa3 <br><br> COC6H6 <br><br> To a mixture of benzonitrile (10 g, 96.97 mmol), concentrated sulfuric acid (10 g) and glacial acetic acid (40 mL) was added, with stirring at 00 C, the diacetate (32, 10 g, 19.43 mmol). The solution was stored for 10 h at room temperture, then poured onto ice (150 mL) and extracted with ether (3 x lOOmL). The ether extracts were washed neutral with water (3 x 100 mL), and <br><br> 59 <br><br> WO 01/58889 PCT/USO 1/04168 <br><br> saturated sodium bicarbonate solution, dried (Na2S04) and evaporated in vacuo. The crude product (21 g) was chromatographed on silica gel eluting with petroleum ether:ether (9:1). The unchanged benzonitrile was first removed. Elution with 5 ether then afforded 11 g (88%) of product that was crystallized from petroleum ether to give analytical sample (33) m.p. 90-92 0 C (lit. 1 94-95° C). <br><br> 2,5,7,8-T p.tramp,thy1-2(4,8,1 9.-trimethy1tridecy1)3.4-10 dihydroquino1in-6(2H)-one (37Q <br><br> 15 <br><br> ^16^33 <br><br> To a solution of the diacetate (33, 13 g, 20.4 mmol) in 60 mL of methanol, solid potassium hydroxide (11 g) was added and the mixture was refluxed under argon for 45 min. After cooling, the solution was diluted with ice-water (100 mL), and extracted under 20 carbon dioxide with ether (3 x 100 mL) yielding 10 g (88%) of crude hydorquinone (34). <br><br> A solution of this product (34, 10 g, 18.1 mmol) in absolute ether (110 mL) was added dropwise with stirring during 20 min. to a suspension of lithium aluminum hydride (20 g, 527 mmol) 25 in absolute ether (110 mL). The mixture was refluxed for 14 h and then hydrolyzed under carbon dioxide by the dropwise addition of methanol (lOmL) followed by 3N hydrochloric acid (30 mL). After extraction with ether (3 x 200 mL), the crude solid benzylamino derivative (35, 8.4 g, 86%) was obtained. <br><br> 60 <br><br> WO 01/58889 PCT/USO1/04168 <br><br> This material (35, 8.4 g, 15.6 mmol) was dissolved in glacial acetc acid (120 mL) and hydrogenated at 50 ° C and atmospheric pressure in the presence of 2 g of palladium o n charcoal (5%) at room temprature for 22 h (or until the hydrogen 5 absorption ceased). After cooling, the catalyst was filtered off, and the solution was diluted with water (50 mL), extracted with ether (3 x 80 mL) and dried (Na2S04). <br><br> To this dry and neutral ether solution of 36 was added fresh silver oxide (2.1 g, 8.93 mmol), and the suspension was 10 stirred for 14 h. After filtration under argon and evaporation, 4 g of the crude iminoquinone (37) was obtained, which was chromatographed on silica gel eluting with hexanes:ether (200:1) to yield 2.4 g (63%) of pure yellow oil. HMRS (cl, m/z): 428.389883 (M + H+, Calc. For C29H49NO 428.389241). <br><br> 15 <br><br> 1 - A 7. a-nr-tocopherol (38.) <br><br> The iminoquinone (37, 1.34 g, 3.133 mmol) in 10 mL of ether was hydorgenated at room temperature and atmospheric pressure in the presence of 190 mg of Lindlar's catalyst. After 20 h, 25 the catalyst was filtered off under argon and the filtrate was evaporated in vacuo to give 810 mg (60%) of crude red oil, which was used in the next step without further pruification. <br><br> 61 <br><br> WO 01/58889 PCT/USO 1/04168 <br><br> l-A7.a-fif-tnrophp.rol-6-yloYyl-arp.tic acid C391 and 1-A/arc -tocopherol -6-v In yyl -m eth yl acetate (40) <br><br> HOgCs^ <br><br> MqO-jC <br><br> c16H33 <br><br> C16H33 <br><br> A solution of 1-Aza-a-tocopherol (38, 800 mg, 1.86 mmol) in DMF (30 mL) was treated with methyl bromoacetate (2.1 10 g, 13.4 mmol) and an excess of powdered NaOH (1.9 g, 48.4 mmol). The resulting yellow suspension was stirred vigorously for 24 h at room temperature. The reaction mixture was acidified with 5N HCl to pH6, and extracted with diethyl ether (3 x 30 mL). The combined ether layers were washed with water (3 x 40 mL), brine (1 x 40 mL), 15 and then dried (Na2S04). The ether solution was concentrated to a yellow oil that was purified by silica gel chromatography eluting with CH2C12:MeOH:Hac (97:2:1) to give 300 mg (33%) of sticky yellow solid (39). 'H-NMR (CDC13, ppm) 4.30 (s, 2H, OCH2), 3.30 (s, 1H, NH), 2.59 (t, 2H, Ar-CH2), 2.18 (s, 3H, Ar-CH3), 2.12 (s, 3H, 20 Ar-CH3), 2.00 (s, 3H, Ar-CH3), 1.80-0.84 (m, 38H, Ar-CH2CH2, NCCH3 and phytyl chain); ,3C-NMR (CDC13, ppm): 12.01, 12.77, 12.96, 19.60, 19.67, 19.72, 21.09, 22.60, 22.71, 24.42, 24.79, 26.56, 27.95, 32.11, 32.74, 37.26, 37.37, 37.42, 37.55, 37.65 39.34, 41.36, 50.92, 69.50 (aliphitic C), 117.69, 118.55, 125.99, 126.39, 25 138.07, 146.05 (Ar-C), 173.18 (COOH); (HRMS (CI,m/z): 488.411055 (M + H+, Calc for C31H53N03 488.410370) and 150 mg (17%) of yellow oil (40). 'H-NMR (CDC13/TMS, ppm) 4.31 (s, 2H, OCH,), 3.85 (S, 3H, C02CH3), 3.32 (S, 1H, NH), 2.61 (T, 2H, Ar-CH2), 2.23 (S, 3H, Ar-CH3), 2.17 (S, 3H, Ar-CH3), 2.01 (S, 3H, Ar-CH3), <br><br> 62 <br><br> WO 01/58889 PCT/US01/04168 <br><br> 1.81-0.86 (M, 38H, Ar-CH2CH2, NCCH3 and phytyl chain); 13C-NMR (CDCI3, ppm) 11.96, 12.67, 12.90, 19.57, 19.63, 19.69, 21.06, 21.95, 22.57, 22.68, 24.38, 24.74, 26.49, 27.91, 32.25, 32.67, 32.70, 37.21, 37.33, 37.53, 39.31, 41.68, 50.43, 51.87, 69.97, 5 (aliphitic C), 116.98, 117.72, 126.01, 126.36, 138.43, 146.18 (Aryl C), 169.83 (G=0); HRMS (CI, m/z): 502.425330 (M + H+, Calc. for C32H55N03 502.426020). <br><br> 1 -Aza-N-mp,thy]-nr.-focopherol-6-y1oYyl-mefhy1 acetate <br><br> 10 (41) <br><br> Me02C^0. <br><br> 'C18H33 <br><br> 15 <br><br> Following the literature procedure 2, a mixture of powdered KOH (500 mg, 7.77 mmol) in DMSO (30 mL) was stirred at. 0°C for 5 min. The methyl acetate (40, 1.3 g, 23.32 mmol) was added, followed immediately by the addition of CH3I (3.3 g, 23.32 20 mmol). Stirring was continued for 0.5 h (0°C) after which the mixture was poured into water. Extracts (CH2C12, 3 x 40 mL) of the final mixture were combined, washed with water (40 mL), brine (40 mL), dried (MgS04) and then evaporated. The resulting yellow oil was chromatographed over silica gel eluting with 5% ETOAC in 25 hexane to give 810 mg (60%) of light yellow oil. 'H-NMR (CDCI3/TMS, ppm) 4.34 (s, 2H, OCH3), 3.84 (s, 3H, C02CH3), 2.55 (t, 2H, Ar-CH,), 2.49 (s, 3H, NCH3), 2.24 (s, 3H, Ar-CH3), 2.20 (s, 3H, Ar-CH3), 2.13 (s, 3H, Ar-CH3), 1.82-0.82 (m, 38H, Ar-CH2CH2, NCCH3 and phytyl chain); 13C-NMR(CDC13, ppm) 11.72, 13.12, 14.27,19.55, <br><br> 63 <br><br> WO 01/58889 PCT/US01/04168 <br><br> 19.68, 20.93, 22.57, 22.66, 22.88, 24.35, 24.74, 25.33, 25.37, 27.90, 32.59, 32.71, 37.23, 37.33, 37.51, 37.63, 38.49, 39.06, 39.29, 51.14, 69.57 (aliphitic C), 125.34, 126.36, 127.42, 130.32, 143.89, 150.33 (Ar-C), 169.73 (C=0). The compound 41 was used in the next step without further purification. <br><br> 1 -A7.a-N-mp.thy1-fY.-top.ophp.ro1-6-y1oxy1- acetic acid (42^ <br><br> H02C&gt; <br><br> 10 /C16H33 <br><br> To the mixture of the ester (41, 740 mg, 14.34 mmol) in ethanol (34 mL) was added dropwise 2N NaOH (9 mL, lOeq. 14.34 15 mmol). After stirring the light yellow solution at room temperature for 4h, the reaction mixture was acidified with 2N HCl to pH6, and then extracted with EtOAc (3 x 100 mL) and the combined organic layers were washed with water (2 x 100 mL), brine (1 x 100 mL), dried (MgS04), and evaporated to give a light yellow oil. The oil 20 was chromatographed over silica gel eluting with 1% HOAc, 30% EtOAc in hexans to give 650 mg (90%) of the product (42) as a colorless oil. 'H-NMR (CDC13/TMS, ppm) 10.43 (s, 1H, C02H), 4.37 (s, 1H, OCH2), 2.51 (t, 2H, Ar-CH2), 2.43 (s, 3H, NCH3), 2.22 (Ar-CH3), 2.12 (s, 3H, Ar-CH3), 1.81-0.80 (m, 38H, Ar-CH2CH2, NCCH3 25 and phytyl chain); I3C-NMR (CDC13, ppm): 11.77, 13.18, 14.28, 19.68, 19.73, 20.80, 20.96, 22.62, 22.72, 23.00, 24.00, 24.41, 24.80, 25.37, 27.80, 32.77, 37.28, 37.30, 37.74, 38.00, 39.00, 39.35, 54.69, 69.17 (aliphitic C), 125.11, 126.55, 127.20, 130.63, <br><br> 64 <br><br> WO 01/58889 PCT/USO 1/04168 <br><br> 142.00, 149.60 (Ar-C), 173.62 (COOH); HRMS (CI, m/z): 502.426327 (M + H+, Calc. For C32H55N03 502.426020). <br><br> 6-f2-4-Dinitrophp,ny1a7.o(9.r51718-tp.tramethy1-2-f4.8.1 2-5 tri m p.thvl tri d ecy 1)-1 9., 3,4-tp.tr a hy d roqn i n ol i n e (43) <br><br> xiN02 <br><br> 10 N 6^33 <br><br> 2,4-dinitrophenylhydrazine (1.95 g, 9.82 mmol) was disolved in 34 mL of hot absolute ethanol and 3.4 mL of concentrated sulfuric acid was carefully added. The hot red 15 solution was cooled to room temperature and the iminoquinone (37, 1.00 g, 2.34 mmol) in 12 mL of absolute ethanol was added with stirring. The mixture was allowed to stand at room temperture for 70 h, then diluted with 120 mL of water and extracted with ether (3 x 100 mL). The combined ether extracts were washed neutral 20 with water (6 x 100 mL), dried (NaS04), and evaporated in vacuo to give 2.9 g of a partly crystalline dark violet mass. This product was chromatographed over silica gel eluting with 5% ether in petroleum ether to yield 1.35 g (95%) of violet crystals (43), m.p. 72-73 &lt;C (lit.1 73-75). HRMS (CI, m/z): 608.417199 (M + H+, Calc. For 25 C35H53N504 608.417581). <br><br> 65 <br><br> WO 01/58889 PCT/USO 1/04168 <br><br> FXAMPTF4 <br><br> Cell Culture Conditions <br><br> All test cell lines were cultured at 37°C in 5% C02 in 5 standard media supplemented with fetal calf serum, using established standard conditions. Plastic adherent cells were disassociated with trypsin, washed, counted, and used directly in experiments. All cells were examined routinely to verify no _ mycoplasma contamination. <br><br> 10 <br><br> FYAMPT.F5 <br><br> Solubility and Dilution of Novel Tocopherol and Tocotrienol <br><br> 15 Compounds <br><br> All compounds were handled as if they were light sensitive (photodegradable). All compounds were initially dissolved in absolute ethanol and subsequently diluted to a final concentration of 0.5% ethanol with the appropriate media. <br><br> 20 <br><br> EXAMPLE 6 <br><br> Determination of Rffective Concentration (KC^ to Tndnce Apoptosis 25 Whereas the parental non-structurally modified forms of tocopherols do not exhibit effective apoptotic properties against a battery of tumor cells, fifteen out of twenty-nine RRR-a-tocopherol compounds and two out of five 1-aza-a-tocopherol analogs, structurally modified via ether linked moieties of different <br><br> 66 <br><br> WO 01/58889 PCT/US01/04168 <br><br> composition and size were extremely effective at inducing tumor cells to undergo apoptosis while having no apoptotic inducing properties on normal cells. Compounds 1, 2, 3, 7, 8, 9, 12, 15, 17, 19, 20, 21, 22, 25, 26, 27, 39, and 42 exhibit effective 5 growth inhibitory (apoptotic inducing) properties specific for human cancer cells from a wide variety of cell lineages, including (i) breast (estrogen responsive Michigan Cancer Foundation human breast cancer cell line number 7, MCF-7 McGuire; non-estrogen responsive M.D. Anderson metastatic breast human cancer cell line, 10 MDA-MB-435; and, estrogen non-responsive M.D. Anderson metastatic human breast cancer cell line, MDA-MB-231); (ii) prostate (androgen responsive human prostate cancer cell line, LnCaP and the androgen non-responsive human prostate cancer cell line, PC-3 and the DU-145 cell line); (iii) promyelocytic leukemia 15 cells (human Promyelocytic Leukemia Cell Line, HL-60), lymphoid cell lines Jurkat and HL-60; (iv) cervical (human cervical cancer cell line, ME-180); (v) ovarian (human ovarian cancer cell line, C-170 cells); (vi) endometrial (human endometrial cancer cell line, RL-95-2 cells); (vii) colon cell lines DLD-1; and (viii) lung cell line A-549. 20 Normal primary breast cells (normal primary early passage human mammary epithelial cells, HMEC) and immortalized, non-tumorigenic mammary cells (Michigan Cancer Foundation immortalized but non-tumorigenic human mammary number 10A cells, MCF-10A) do not undergo apoptosis when cultured with the 25 above pharmacodynamically designed forms of tocopherol. <br><br> The effective therapeutic dose of novel reagents for controlling cancer growth is referred to as the growth inhibitory concentration (IC50) or effective concentration (EC50) that blocks 50% cancer growth via DNA synthesis inhibition, cell cycle blockage <br><br> 67 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> and/or cell death. The apoptotic ECS0 for a battery of test cancer cells for the twenty-nine novel RRR-a-tocopherol compounds and two of the five 1-aza-a-tocopherol analogues of this invention are presented in Tables 1 and 2. <br><br> 68 <br><br> TABLE 1 <br><br> Apoptosis Induced by Novel Tocopherol Compounds (ec^ranqe jig/ml) <br><br> Cell Type <br><br> VES <br><br> 1 <br><br> 2 <br><br> 3 <br><br> 4 <br><br> 5 <br><br> 6 <br><br> 7 <br><br> 8 <br><br> 9 <br><br> 1 0 <br><br> 11 <br><br> 12 <br><br> 13 <br><br> 14 <br><br> 15 <br><br> Breast Cancer <br><br> HMEC <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> NT <br><br> MCF-10A <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> NT <br><br> MDA-MB-435 <br><br> 5-10 <br><br> 5-10 <br><br> 10-20 <br><br> 5-10 <br><br> N <br><br> N <br><br> N <br><br> 5-10 <br><br> 5-10 <br><br> 5-10 <br><br> N <br><br> N <br><br> 5-10 <br><br> N <br><br> N <br><br> 20-30 <br><br> MDA-MB-231 <br><br> 5-10 <br><br> 5-10 <br><br> 10-20 <br><br> 5-10 <br><br> N <br><br> N <br><br> N <br><br> 5-10 <br><br> 5-10 <br><br> 5-10 <br><br> N <br><br> N <br><br> 5-10 <br><br> N <br><br> N <br><br> 20-30 <br><br> MCF-7 <br><br> 10-20 <br><br> 5-10 <br><br> 20-30 <br><br> 20-30 <br><br> N <br><br> N <br><br> N <br><br> 10-20 <br><br> 10-20 <br><br> 10-20 <br><br> N <br><br> N <br><br> 5-10 <br><br> N <br><br> N <br><br> 20-30 <br><br> T47D <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Cervical <br><br> MH-180 <br><br> 10-20 <br><br> 1-5 <br><br> 5-10 <br><br> 10-20 <br><br> N <br><br> N <br><br> N <br><br> 5-10 <br><br> 5-10 <br><br> 5-10 <br><br> N <br><br> N <br><br> 5-10 <br><br> N <br><br> N <br><br> N <br><br> Ovarian <br><br> C-170 <br><br> N <br><br> 10-20 <br><br> 10-20 <br><br> 10-20 <br><br> N <br><br> N <br><br> N <br><br> 10-20 <br><br> 10-20 <br><br> N <br><br> N <br><br> N <br><br> 10-20 <br><br> N <br><br> N <br><br> N <br><br> Endomerial <br><br> RL-95-2 <br><br> 10-20 <br><br> 10-20 <br><br> 10-209 <br><br> 10-20 <br><br> N <br><br> N <br><br> N <br><br> 5-10 <br><br> 1-5 <br><br> 5-10 <br><br> N <br><br> N <br><br> 5-10 <br><br> N <br><br> N <br><br> N <br><br> Prostate <br><br> PREC <br><br> N <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> LnCaP <br><br> 5-10 <br><br> 5-10 <br><br> 5-10 <br><br> 5-10 <br><br> N <br><br> N <br><br> N <br><br> 2.5-5 <br><br> 5-10 <br><br> 5-10 <br><br> N <br><br> N <br><br> &gt;20-30 <br><br> NT <br><br> N <br><br> NT <br><br> PC-3 <br><br> 10-20 <br><br> 5-10 <br><br> 5-10 <br><br> 5-10 <br><br> N <br><br> N <br><br> N <br><br> 5-10 <br><br> 5-10 <br><br> 5-10 <br><br> NT <br><br> N <br><br> 10-20 <br><br> N <br><br> N <br><br> NT <br><br> DU-145 <br><br> 10-20 <br><br> 5-10 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Colon <br><br> HT-29 <br><br> 5-10 <br><br> 10-20 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> DLD-1 <br><br> 10-20 <br><br> 10-20 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Luna <br><br> A-549 <br><br> 20-30 <br><br> 10-20 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> LvmDhoid Cells <br><br> Myeloma <br><br> 10-20 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Raii <br><br> 10-20 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Ramos <br><br> 10-20 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Jurkat <br><br> 10-20 <br><br> 10-20 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> HL-60 <br><br> 10-20 <br><br> 5-10 <br><br> 10-20 <br><br> 10-20 <br><br> N <br><br> N <br><br> N <br><br> 5-10 <br><br> 10-20 <br><br> 10-20 <br><br> N <br><br> N <br><br> 20-30 <br><br> N <br><br> N <br><br> NT <br><br> EC*,50 pg/ml of tocopherol coinpunds I -29 inducing 509S- apoptosis; N = No apoptosis when treated for 2 days with I -60 EC,,, Hg/rol or tocopherol compounds I -29; NT = Not tented; * = compound* exhibiting toxicity. <br><br> TABLE 2 <br><br> Apoptosis Induced by Novel Tocopherol Compounds (ec^range fiq/mh <br><br> Cell Type <br><br> 1 6 <br><br> 1 7 <br><br> 1 8 <br><br> 1 9 <br><br> 20 <br><br> 21 <br><br> 22 <br><br> 23 <br><br> 24 <br><br> 25 <br><br> 26 <br><br> 27 <br><br> 28 <br><br> 29 <br><br> 39 <br><br> 42 <br><br> 43 <br><br> Breast Cancer <br><br> HMEC <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> MCF-10A <br><br> NT <br><br> N <br><br> NT <br><br> N <br><br> N <br><br> N <br><br> NT <br><br> N <br><br> NT <br><br> N <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> MDA-MB-435 <br><br> N <br><br> NT <br><br> N <br><br> 10-20 <br><br> 10-20 <br><br> N <br><br> NT <br><br> N <br><br> N <br><br> N <br><br> 20-40 <br><br> NT <br><br> NT <br><br> NT <br><br> 10-20 <br><br> 10-20 <br><br> PPT. <br><br> MDA-MB-231 <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> MCF-7 <br><br> N <br><br> 10-20 <br><br> N <br><br> 10-20 <br><br> 5-10 <br><br> N <br><br> 15-20 <br><br> N <br><br> N <br><br> N <br><br> N <br><br> 10-20 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> T47D <br><br> NT <br><br> 10-20 <br><br> NT <br><br> N <br><br> 5-10 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Cervical <br><br> ME-180 <br><br> NT <br><br> 20-30 <br><br> N <br><br> 1-5 <br><br> 1-5 <br><br> 1-5 <br><br> NT <br><br> NT <br><br> NT <br><br> N <br><br> * <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Ovarian <br><br> C-170 <br><br> NT <br><br> 20-30 <br><br> N <br><br> 1-5 <br><br> * <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> N <br><br> 20-30 <br><br> NT NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Endomerial <br><br> RL-95-2 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> N <br><br> 20-30 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Prostate <br><br> PREC <br><br> NT <br><br> NT <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> LnCaP <br><br> NT <br><br> 10-20 <br><br> NT <br><br> 5-10 <br><br> 5-10* <br><br> N <br><br> NT <br><br> NT <br><br> N <br><br> N <br><br> 10-20 <br><br> 10-20 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> PC-3 <br><br> NT <br><br> NT <br><br> NT <br><br> N <br><br> 5-10* <br><br> N <br><br> NT <br><br> N <br><br> N <br><br> 20-30 <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> DU-145 <br><br> NT <br><br> NT <br><br> NT <br><br> 5-10 <br><br> 5-10" <br><br> N <br><br> NT <br><br> N <br><br> N <br><br> 20-30 <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Colon <br><br> HT-29 <br><br> NT <br><br> N <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> N <br><br> NT <br><br> N <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> DLD-1 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> N <br><br> N <br><br> 20-40 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Luna <br><br> A-549 <br><br> NT <br><br> N <br><br> N <br><br> 20-30 <br><br> 20-30 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> LvmDhoid Cells <br><br> Myeloma <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Raji <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Ramos <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> Jurkat <br><br> NT <br><br> 10-20 <br><br> N <br><br> 10-20 <br><br> 10-20 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> 20-30 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> HL-60 <br><br> NT <br><br> 10-20 <br><br> N <br><br> 10-20 <br><br> 1 0 <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> N <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> NT <br><br> EC*,50 Hg/ml of tocopherol compunds 1-29 inducing 50% apoptosis; N = No apoptosis when treated for 2 days with 1-60 ECj„ |_ig/ml of tocopherol compounds 1-29; NT = Not texted; * = compounds exhibiting toxicity. <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> FXAMPT.F7 <br><br> Rioassay for Apoptosis <br><br> Cells were cultured at 1.5 x 105 cells/well in 12 well 5 plates. Cells were allowed to adhere overnight, then incubated with novel test compounds at 0.01, 0.1, 1, 5, 10 &amp; 20 jig/mL for 1, 2 and 3 days. After treatment, cells (floating + trypsin released adherent cells) were pelleted, washed and stained with 2 |xg/ml DAPI (4',6-diamidine-2'-phenylindole dihydrochloride) in 100% methanol for 10 15 minutes at 37°C and/or TUNEL stained, then viewed using a Zeiss ICM 405 microscope. Cells whose nucleus contained clearly condensed or fragmented chromatin were scored as apoptotic. Data are presented as percent cells undergoing apoptosis. <br><br> 15 <br><br> FXAMPIFS <br><br> Rioassay for T)NA synthesis arrest <br><br> To assay DNA synthesis, all cells were used at 2.5 x <br><br> 20 10^/ml. Cells were treated with each of the compounds 1-29 (Tables 1 and 2) at concentrations of 0.01, 0.1, 1, 5, 10 and 20 fig/mL and 200 jxl of each treatment group were plated in quadruplicate in a 96 well culture plate (Corning, Corning NY). Experiments were done in duplicate, one plate used for viability 25 testing and the other plate for examination of ^H-TdR uptake to monitor DNA synthesis. Plates were cultured for 48 hours at 37°C, 5% CO2. Eight hours prior to the end of incubation, ^H-TdR was added to one of the duplicate plates and incubation continued for 8 hours. The cells were then harvested (trypsinization was required <br><br> 71 <br><br> WO 01/58889 PCT/USO 1/04168 <br><br> to harvest adherent cells), and isotope uptake was determined as counts per minute (cpm). For viability studies, at the end of the incubation, the cells were removed from the wells and viability checked by the Trypan Blue Exclusion method. Percent viability and 5 percent DNA synthesis in comparison to untreated or vehicle treated cells of each treatment group were calculated. <br><br> EXAMPLE 9 <br><br> 10 <br><br> Bioassay for Cell Cycle Arrest <br><br> The cells were cultured with novel test agents for 2-3 days, fixed in 95% ethanol and stained with propidium iodide overnight. DNA content was determined using a Coulter Epics Elite 15 Flow Cytometer with an argon laser setting of 488 nm. Cell size was measured simultaneously, and data were analyzed as to percent cells in each cell cycle phase using the Coulter Multicycle Program. <br><br> 20 EXAMPLE 10 <br><br> Bioassay for Cellular Differentiation <br><br> To determine if the novel compounds were inducing cellular differentiation, the cells were cultured on cover slips, fixed 25 in 95% ethanol and stained with a lipid specific stain for detection of milk lipids. Additionally, cells were examined by immunohistology and by Western analyses for presence of milk protein casein, using polyclonal antibodies produced in the lab. <br><br> 72 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> EXAMPLE 11 <br><br> DNA Synthesis Arrest Kffccts <br><br> The cells were cultured for 48 hours, pulsed 8 hours 5 with tritiated thymidine, harvested and counted. Data are presented as counts per minute. Verification of DNA synthesis arrest is determined by reduced tritiated thymidine uptake by cells treated with test compounds. Further verification of DNA synthesis arrest is determined by propidium iodide staining and standard cell cycle 10 analyses. <br><br> EXAMPLE 12 <br><br> Mechanisms of Induction of Apoptosis <br><br> The mechanism of induction of apoptosis by these compounds appears to involve three distinct apoptotic signaling pathways; namely, activation of latent transforming growth factor-beta (TGF-P), activation of the Fas/Fas ligand signaling pathways, and signaling by the stress kinase (c-Jun N-terminal Kinase) pathway. <br><br> TGF-ps are potent growth inhibitory molecules that are known to inhibit cell growth by inhibition of DNA synthesis arrest and by induction of apoptosis. TGF-ps are involved only in induction of the apoptotic pathway, i.e., there is no evidence curently that the TGF-Ps effect DNA synthesis arrest; however, this possibility has not been completely ruled out. TGF-ps are made and secreted by cells in a latent non-active form. To be effective as tumor growth inhibitors, the latent TGF-Ps must be activated by <br><br> 15 <br><br> 20 <br><br> 25 <br><br> 73 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> induction of cell surface proteins that provide a proper structure for processing and activating proteases that cut the latent protein and release the active TGF~p. <br><br> The compounds of the present invention are shown t o 5 activate proteases such as cathepsin D family proteases, and upregulate the mannose-6-phosphate receptor which binds inactive TGF-P and permits activation via proteases. Active TGF-P signals via cell membrane TGF-P receptors I and II to activate down stream kinases referred to as stress kinases or c-Jun N-terminal Kinases 10 (JNK) which phosphorylate and activate transcription factors c-Jun, ATF-2 and Elk-1. Prolonged activation of transcription factor c-Jun causes tumor cells to undergo apoptosis. These transcription factors, acting as homodimers or heterodimers with a multitude of transcription factor partners activate proapoptotic genes and/or 15 downregulate antiapoptotic genes leading to DNA fragmentation. The compounds of the present invention do not generate an antiproliferative outcome to TGF-p signaling in normal non-tumor cells. <br><br> A second apoptotic inducing mechanism called the Fas/Fas ligand apoptotic signaling pathway is activated by the novel 20 compounds of the present invention. Activated Fas/Fas ligand signaling may lead to rapid cell death by apoptosis. Thus, for tumor cells to escape death by Fas/Fas ligand, they must inactivate this most important apoptotic pathway. The mechanism for inactivation of the Fas/Fas ligand signaling pathway by tumor cells varies; 25 however, many tumor cells down regulate the expression of Fas receptor and Fas ligand on their membranes. <br><br> Most important, R,R,R-2-(2,5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)chroman-6-yloxy)acetic acid (1) has been shown to induce Fas/Fas ligand resistant tumor cells to become Fas/Fas <br><br> 74 <br><br> WO 01/58889 <br><br> PCT/USO 1/04168 <br><br> ligand sensitive. Compound 1 also has the ability to enhance the expression of Fas ligand on the membrane of LNCaP prostate cells. Studies show that Fas signaling resistant human breast cancer cells retain the Fas receptor in their cytoplasm, but when cultured with 5 compound 1, the Fas receptor is transported from the cytoplasm to the membrane; thereby rendering the cells Fas signaling sensitive. Furthermore, this compound is synergistic in anti-Fas triggered apoptosis in that greater amounts of cell killing is obtained with both human breast and prostate cancer cells when co-treated versus 10 when treated separately. The ability of compound 1 to convert Fas signaling resistant tumor cells to Fas signaling sensitive tumor cells and to exhibit synergistic killing effects provides an extremely important mechanism for destruction of tumor cells both by the host immune surveillence system as well as by pharmaceutical 15 intervention. The compounds of the present invention do not activate the Fas signaling pathway of normal non-tumor cells. <br><br> These compounds activate the JNK kinase signaling pathway, perhaps by TGF-P and Fas/Fas ligand signaling. Prolonged activation of JNK results in prolonged activation of c-Jun and ATF-2 20 transcription factors, which are postulated to play a role in expression or repression of proapoptotic and antiapoptotic genes, respectively. <br><br> 25 <br><br> EXAMPLE 13 <br><br> Mechanism of Induction of DNA Synthesis Arrest, Cell Cycle Arrest and Cellular Differentiation <br><br> The mechanisms of growth inhibition by DNA synthesis arrest, cell cycle arrest and by induction of cellular differentiation <br><br> 75 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> have not been characterized as fully as the mechanism of growth inhibition by apoptosis. Studies show that the compounds of the present invention have profound effects on the cell cycle, inducing DNA synthesis arrest of approximately 95% of the tumor cells within 5 24 hours of treatment. Tumor cells cultured with the compounds disclosed herein are growth inhibited in the G1 cell cycle phase, undergo morphological changes and express milk lipids, an indication that the cell cycle blocked cells have undergone differentiation. P21, a gene known to be an inhibitor of entrance of 10 cells from the G1 cell cycle phase to the S phase of the cell cycle, and the mRNA, as well as the protein of P21 gene, is up-regulated by treatment of MDA-MB-435 human breast cancer cells with compound 1. <br><br> 15 <br><br> EXAMPLE 14 <br><br> Tn Vivo Potp.nt.ia1 for Human Cancer Cells <br><br> The present invention has potential for use a s 20 therapeutic agents. In vivo studies of tumor growth and metastasis of human tumor cells either ectopically or orthotopic ally transplanted into immune compromised animals, such as nude mice, or in vivo studies employing well recognized animal models are conducted. Inhibition of growth of human tumor cells 25 transplanted into immune compromised mice provides pre-clinical data for clinical trials. In vivo studies include two human tumor cell models, the metastatic non-estrogen responsive MDA-MB-435 breast cancer model, and the androgen non-responsive PC-3 prostate cancer model. <br><br> 76 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> MDA-MR-4^5 Breast Cancer Model: <br><br> Pathogen free MDA-MB-435 human breast cancer cells stably transfected with a marker protein (fluorescent green protein) are grown as a solid tumor in immune compromised nude or SCID 5 mice. The tumors are removed, and 1 mm sections of equal size are orthotopically transplanted into the mammary fat pad or ectopically transplanted into the hind flank of female nude mice. Tumor growth, metastasis, and death of the animals are determined. Tumor growth is measured by caliper evaluations of tumor size. At 10 the time of sacrifice, tumors are removed, measured for size, and used for histochemical examination. Organs such as spleen, lymph nodes, lungs, and bone marrow, are examined for metastatic MDA-MB-435 cells by histochemical staining of tissue sections for expression of the marker fluorescent green protein. <br><br> 15 <br><br> PC-3 Prostate Cancer Model <br><br> Pathogen free PC-3 human prostate cancer cells stably transfected with a marker protein (fluorescent green protein) are grown as a solid tumor in nude mice. The tumors are removed, and 20 1 mm sections of equal size are ectopically transplanted into the hind flank of male nude mice. Tumor growth, metastasis, and death of the animals are determined. At the time of sacrifice, tumors are removed, measured for size, and used for histochemical examination. Organs such as spleen, lymph nodes, lungs, bone 25 marrow, are examined for metastatic PC-3 cells by histochemical staining of tissues for expression of the marker fluorescent green protein. <br><br> 77 <br><br> WO 01/58889 <br><br> PCT/USO 1/04168 <br><br> SVin Cancer Animal Model <br><br> Skin cancer is induced in SENCAR and SKH-1 hairless mice by ultraviolet irradiation and chemical (DMBA) treatments. In addition, mice specifically expressing the oncogene Her-2/neu in 5 skin basal cells that spontaneously develop skin cancer are used. The compounds disclosed herein are topically applied to the skin daily, before and after skin cancer initiation, and development of skin papilloma formation is assessed. Control mice are treated identically except that they receive vehicle treatments topically 10 applied to their skin. The efficacy of these compounds in treating papilloma's as well as their ability to affect malignant conversion when supplied prior to premalignant progression is monitored. <br><br> 15 EXAMPLE 15 <br><br> Supplementation with Novel Compounds <br><br> Prior to initiation of the in vivo experiments, the compounds of this invention that exhibit the greatest amount of 20 tumor cell killing are adminstered to nude, SCID, transgene, and other mice at varing levels to establish the highest level of compound that can be administered safely without adverse effects. The compounds are administered in a model-appropriate manner; e.g., orally, injections, including injections directly into the target 25 organ, or topically. After establishing the highest level of the compounds that can be tolerated and effective administration routes, the novel compounds are administered to the mice on a daily basis, and tumor growth and progression is determined as described above. <br><br> 78 <br><br> WO 01/58889 PCT/US01/04168 <br><br> EXAMPLE 16 <br><br> Establishing Maximum Tolerated Dose (MTD) of Compound 1 <br><br> Prior to conducting the preclinical chemotherapeutic 5 studies, compound 1 is assayed in non-tumor Balb/c female mice in an effort to establish the maximum tolerated dose. Compound 1 is dissolved in 100% ethanol to establish a stock solution (2 grams of compound 1 in 5 mis of 100% ethanol) and diluted in peanut oil (non vitamin E supplemented) to yield 20, 10, and 5 mg/0.1 ml. 10 RRR-a-tocopherol succinate (VES; the succinate moiety is attached to RRR-a-tocopherol via an ester linkage) was solubilized and diluted as described above to yield 20 mg/0.1 ml/gavage, serving as an additional control. A total of 30 Balb/c female mice were placed into 6 groups (5 mice/group) and supplemented by gavage with 15 compound 1 daily for a total of 23 days. Group 1 mice were non treated and served as control; group 2 mice were the vehicle control group and received 0.1ml of vehicle (ethanol + peanut oil equivalent to treatment groups)/mouse/day for a total of 23 days, group 3 mice received 20 mg/ml of VES daily for a total of 23 days. 20 Treatment groups 3, 4 and 5 were gavaged daily for 23 days with 5, 10, and 20 mgs of compound 1. Compound 1 at 5, 10, and 20 mg/day, administered daily by gavage to mice in groups 3, 4, and 5, respectively, for 23 days is tolerated well by the mice. Weights are determined weekly and there is no significant difference in the 25 weights of the mice receiving compound 1 at 5, 10, and 20 mg/day when compared with the weights of the VES treated, untreated and vehicle treated control mice (Figure 8, data is for days 11 and 2 3 days treatment). Mice remained active and showed no signs of <br><br> 79 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> toxicity. At the completion of the MTD studies, mice were sacrificed and no evidence of toxicity was observed upon autopsy. <br><br> 5 EXAMPLE 17 <br><br> Chemotherapeutic effectiveness of compound 1 <br><br> The maximum tolerated dose studies show that compound 1 administered at the highest level (20 mg/day/23 days) 10 is not toxic; however, due to the non-toxic levels used, an maximum tolerated dose was not established. For the preclinical chemotherapeutic studies, in the absence of an maximum tolerated dose, the mice are supplemented with 30 and 20 mgs/O.lml by gavage daily for 21 days. Compound 1 is dissolved in 100% ethanol 15 at 2 grams of compound l/5mls of ethanol, and diluted in peanut oil (nonsupplemented vitamin E) to yield 30 and 20 mgs of compound 1 per 0.1ml. Vehicle control consisted of a mixture of ethanol and peanut oil equivalents. The preclinical chemotherapeutic studies are conducted with compound 1, utilizing 20 human MDA-MB-435 human breast cancer cells, human DU-145 prostate cancer cells, and human HT-20 colon tumor cells transplanted into Balb/c immune compromised nude mice. <br><br> A total of 40 immune compromised nude mice are utilized for testing each test compound (two treatment groups at 2 0 25 and 30 mg/day) in each tumor cell type. Four experimental treatment groups of 10 mice each (vehicle/0.lml/gavage daily for 21 days, compound 1 at 30 mgs of compound #1/0. lml gavage/daily for 21 days, and compound 1 at 20 mgs/of compound 1/0. lml gavage daily for 21 days, for each tumor type are used. An <br><br> 80 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> established dosage of effective chemotherapeutic drugs are used in the xenograft studies. Taxol at 20/mg/kg, administered intraperitioneally daily for 5 days, is used for a positive control for human MDA-MB-435 breast cancer xenografts; Mitoxantrone at 1 5 mg/kg, administered daily intravenously for 5 days, is used as the positive control for human DU-145 prostate cancer cells; and 5 Fluorouracil (5 FU) at 30 mg/kg, administered daily for 5 days, is used for a positive control for human HT-29 colon cancer cells. Tumor cells are transplanted subcutaneously into the left flank and 10 permitted to grow to approximately 70 mg in size, test compound treatments by daily gavage are initiated, and continue for a total of 21 days. Mice are monitored daily for tumor size by caliper measurements. Mice are treated for 21 days; however, the mice are sacrificed when the tumors of the vehicle control reached 500 15 mg in weight. At the completion of the protocol, tumors are excised and tumor size established by weight. At the time of sacrifice, serum, tumor, and heart muscle are taken from animals in the two compound 1 test groups in order to establish levels of compound 1 via HPLC analyses (data not included). The 20 chemotherapeutic effectiveness of compound 1 administered daily at 30 and 20 mgs/day is determined by comparing the tumor size of the two treatment groups with the vehicle control group for each of the three tumor types, and by comparing the size of the tumor in the two treatment groups with the tumor group from the positive 25 control. Treatment is for 21 days, experiments are continued until mean weight of tumor for vehicle control reaches 500 mgs. <br><br> The in vivo preclinical chemotherapeutic data are depicted in Tables 3 and 4 and Figure 9A-C and are presented for 21-22 days of treatment. Treatment of nude mice transplanted with <br><br> 81 <br><br> WO 01/58889 <br><br> PCT/US01/04168 <br><br> human MDA-MB-435 breast cancer cells with compound 1 for 21 days at 20 and 30 mg/day by gavage reduced the mean tumor weight (mg) of the transplanted tumor cells by 20.7 and 44.6%, respectively when compared to the mean tumor weight of the 5 vehicle control group at day 21 after treatment. The positive control, Taxol, reduces the mean tumor growth, when compared to mean tumor growth of vehicle control group, by 90.3% (Table 3, 4 &amp; Figure 9A). Data are converted to percent mean tumor growth in relation to weight gain at the beginning of treatment (day 1) and are 10 depicted in Table 4. Treatment of nude mice transplanted with human DU-145 prostate cancer cells with compound 1 for 21 days at 20 and 30 mg/day by gavage reduces the mean tumor weight (mg) of the transplanted tumor cells by 31.5 and 24.8%, respectively, when compared to the mean tumor weight of the 15 vehicle control group at day 22 (Table 3, 4 &amp; Figure 9B). The positive control, mitoxantrone reduces the mean tumor growth, when compared to mean tumor growth of vehicle control group, by 27.9%. Treatment of immune compromised nude mice transplanted with human HT-29 human colon cancer cells with 20 compound 1 for 21 days at 20 and 30 mg/day by gavage reduces the mean tumor weight (mg) of the transplanted tumor cells by 33.3 and 34.5 %, respectively when compared to the mean tumor weight of the vehicle control group at day 22 (Table 3, 4 &amp; Figure 9C). The positive control, 5 Fluorouracil, reduces the mean tumor growth, 25 when compared to mean tumor growth of vehicle control group, by 31.4%. <br><br> 82 <br><br> WO 01/58889 <br><br> PCT/USO 1/04168 <br><br> TABLE 3 <br><br> MDA-MB-435 Human Breast Cancer Cells Transplanted in to Nnde <br><br> Mice <br><br> Mean tumor Weights (mg) Following Treatments <br><br> 5 <br><br> Treatments <br><br> 1 <br><br> 4 <br><br> 7 <br><br> 1 n <br><br> 1 4 <br><br> 17 <br><br> 91 <br><br> Vehicle <br><br> 70.1 <br><br> 98.2 <br><br> 119.5 <br><br> 153.2 <br><br> 172.8 <br><br> 203.7 <br><br> 226.1 <br><br> #1 (20 mg) <br><br> 70.1 <br><br> 85.8 <br><br> 123.9 <br><br> 127.9 <br><br> 147.2 <br><br> 143.8 <br><br> 179.4 <br><br> #1 (30 mg) <br><br> 68.1 <br><br> 85.7 <br><br> 97.3 <br><br> 98.4 <br><br> 126.6 <br><br> 118.7 <br><br> 125.2 <br><br> Taxol <br><br> 70.7 <br><br> 71.5 <br><br> 38.2 <br><br> 11.7 <br><br> 5.7 <br><br> 3.6 <br><br> 6.8 <br><br> 10 <br><br> DU-145 Human Prostate Cancer Cells Transplanted into Nnde Mice <br><br> Mean tumor Weights <br><br> (mg) Following Treatments <br><br> Treatments <br><br> 1 <br><br> 5 <br><br> 8 <br><br> 1 9. <br><br> 1 5 <br><br> 19 <br><br> 9 2 <br><br> Vehicle <br><br> 66.5 <br><br> 111.4 <br><br> 167.1 <br><br> 258.5 <br><br> 351.1 <br><br> 458.3 <br><br> 540.9 <br><br> #1 (20 mg) <br><br> 66.5 <br><br> 103.5 <br><br> 137.9 <br><br> 208 <br><br> 254.8 <br><br> 362.2 <br><br> 370.5 <br><br> 15 <br><br> #1 (30 mg) <br><br> 67.8 <br><br> 102.5 <br><br> 131.3 <br><br> 241.2 <br><br> 256.8 <br><br> 379.9 <br><br> 407 <br><br> Mitoxantrone 66.8 <br><br> 107.2 <br><br> 141.1 <br><br> 185.4 <br><br> 205.2 <br><br> 325.3 <br><br> 389.4 <br><br> HT-99 Human <br><br> Colon Cancer Cells Transplanted into Nnde Mice <br><br> Mean tumor Weights <br><br> (mg) Following <br><br> Treatments <br><br> Treatments <br><br> 1 <br><br> 5 <br><br> 8 <br><br> 1 9. <br><br> 1 5 <br><br> 1 9 <br><br> 22 <br><br> 20 <br><br> Vehicle <br><br> 80.3 <br><br> 91.7 <br><br> 133.2 <br><br> 185.6 <br><br> 204.8 <br><br> 260.1 <br><br> 372.2 <br><br> #1 (20 mg) <br><br> 59.5 <br><br> 84.0 <br><br> 97.0 <br><br> 124.6 <br><br> 147.3 <br><br> 198.0 <br><br> 248.2 <br><br> #1 (30 mg) <br><br> 40.0 <br><br> 83.0 <br><br> 96.7 <br><br> 129.3 <br><br> 144.1 <br><br> 176.1 <br><br> 243.8 <br><br> 5-FU <br><br> 59.3 <br><br> 87.5 <br><br> 105.8 <br><br> 145.8 <br><br> 141.6 <br><br> 212.3 <br><br> 255.5 <br><br> 83 <br><br> WO 01/58889 <br><br> PCT/USO 1/04168 <br><br> Table 4: <br><br> Percent mean tumor weight gain (mg) following treatments MDA-MB-435 Human Breast Cancer Cells <br><br> Percent Mean tumor Weight Gain (mg) Following Treatments1 <br><br> 5 Treatments ] 4 1 ; 1.0 IA 12 2JL <br><br> Vehicle 100 140.1 170.5 218.5 246.5 290.6 322.5 <br><br> #1 (20 mg) 100 122.4 176.7 182.5 210.0 205.1 255.9 #1 (30 mg) 100 125.8 142.9 144.5 185.9 174.3 178.6 Taxol 100 101.1 -46.0 -83.7 -91.8 -99.5 -90.3 <br><br> 10 DU-145 Human Prostate Cancer Cells <br><br> Percent Mean tumor Weight Gain (mg) Following Treatments <br><br> Treatments L 5 8 1.2 L5 1.2 22 <br><br> Vehicle 100 167.5 251.3 388.7 528.0 458.3 689.2 <br><br> #1 (20 mg) 100 155.6 207.4 312.8 383.2 362.2 544.7 15 #1 (30 mg) 100 151.2 193.7 355.8 378.8 379.9 560.3 Mitoxantrone 100 160.5 211.2 277.5 307.2 487.0 582.9 HT-29 Human Colon Cancer Cells <br><br> Percent Mean tumor Weight Gain (mg) Following Treatments <br><br> Treatments ] 5 8 L2 L5 L9 12 <br><br> 20 Vehicle 100 152.1 220.9 307.8 339.6 431.3 617.2 #1 (20 mg) 100 141.2 163.0 209.4 247.6 332.8 417.1 #1 (30 mg) 100 139.5 162.5 217.3 242.2 296.0 409.7 5-FU 100 147.6 178.4 245.9 247.2 358.0 430.0 <br><br> 'Percent Mean Tumor Weight Gain (mg) following treatment was determined 25 by dividing the mean tumor weight at various time periods following treatment by the mean tumor weight at day 1 within the same treatment group and multiplying by 100. <br><br> 30 <br><br> 84 <br><br> WO 01/58889 PCT/US01/04168 <br><br> EXAMPLE 18 <br><br> Preparation of Stock Solution, Vehicle and Compound 1 Dilutions <br><br> Compound 1 was prepared weekly, and stored at 4°C. 5 The preparation procedures are as follows: <br><br> Stock Solution of Compound 1: <br><br> Dissolve 2 grams of compound 1 in 5 mis of 100% ethanol (ETOH) and vortex at 37°C. This is the maximum amount of compound 1 that will go into solution. <br><br> 10 For the following dilutions, dry compound 1 is added to yield the appropriate levels of compound 1 while keeping the ethanol levels equal in the two experimental groups and the vehicle control group. <br><br> Compound 1 at 30 mg/0 1ml gavagp./monse: <br><br> 15 Combine 1 ml of compound 1 stock solution, 3 <br><br> mis of vitamin E depleted peanut oil and 800 mg of compound # 1 (dry) and vortex at 37°C. <br><br> Compound 1 at 20 mg/0.1 ml gavage/monse: <br><br> Combine 1 ml of compound 1 stock solution, 3 mis of 20 vitamin E depleted peanut oil and 400 mg of compound #1 (dry) and vortex at 37°C approximate 2 h until in solution. <br><br> Compound 1 at 10 mg/0.1 ml gavage/mouse: <br><br> Combine 1 ml of compound 1 stock solution and 3 mis of vitamin E depleted peanut oil. <br><br> 2:5 Vehicle: <br><br> Combine 1 ml ETOH 3 mis of vitamin E depleted peanut oil. <br><br> 85 <br><br> WO 01/58889 PCT/US01/04168 <br><br> EXAMPLE 19 <br><br> ChemQpreyentive properties of compound 1 in an ACT rat cancer model. <br><br> 5 Compound 1 is used in vivo to treat transplanted human breast, prostate, and colon tumors transplanted in immune compromised nude mice. The chemopreventive effectiveness of compound 1 in vivo against human breast cancer is shown in an estrogen cancer initiated ACI rat breast cancer model. 10 Approximately 90% of rats implanted with estrogen pellets develop breast cancer within 6 months after estrogen implantation. <br><br> Compound 1 is dissolved in 100% ethanol and is diluted to the appropriate dosage using vitamin E depleted peanut oil. The maximum tolerated dose (MTD, maximum dose of compound that 15 can be administered without adverse affects) is determined as described in Examples 14 and 15. Compound 1 is administered at the maximum tolerated dose and 50% of the maximum tolerated dose. ACI rats at 4 weeks of age are subpannicularly implanted with estrogen pellets in the shoulder region. Compound 1 at maximum 20 tolerated dose and 50% of the maximum tolerated dose is administered by gavage Breast tumors are detected in the control group at approximately 100 days following estrogen implantation. Ninety percent of the control rats develop breast cancer within 6 months after estrogen implantation. Tumor bearing animals from 25 control and treatment groups are sacrificed at various time intervals after treatment initiation, and mammary tissue is examined for obvious tumors, and farther examined by histological analyses. <br><br> 86 <br><br> WO 01/58889 PCT/US01/04168 <br><br> The following references are cited herein. <br><br> 1. Colowisk, Sidney, P. and Kaplan, Nathan, O. Methods in Enzymology, Vol. XVIII, Vitamins and Coenzymes, Part C. Edited by Donald B. McCormick and Lemuel D. Wright. Section XH, <br><br> 5 PP. 335. <br><br> 2. Dhar, A., Liu, S., Klucik, J., Berlin, K.D., Madler, M.M., Lu, S., Ivey, R.T., Zacheis, D., Brwon, C.W., Nelson, E.C., Birchbichler, P.J., Benbrook, D.M. Synthesis, Structure-Activity Relationships, and RARy-Ligand Interactions of Nitrogen <br><br> 10 Heteroarotinoids. /. Med. Chem. 42, pp. 3602-3614 (1999). <br><br> Any patents or publications mentioned in this specification are indicative of the levels of those skilled in the art to which the invention pertains. Further, these patents and publications are incorporated by reference herein to the same 15 extent as if each individual publication was specifically and individually indicated to be incorporated by reference. <br><br> One skilled in the art will readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent 20 therein. The present examples along with the methods, procedures, treatments, molecules, and specific compounds described herein are presently representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those 25 skilled in the art which are encompassed within the spirit of the invention as defined by the scope of the claims. <br><br> 87 <br><br></p> </div>

Claims (1)

  1. <div class="application article clearfix printTableText" id="claims"> <p lang="en"> WHAT WE CLAIM IS:<br><br> 1. A compound having a structural formula<br><br> R ■<br><br> R1<br><br> &gt;5<br><br> wherein X is oxygen, nitrogen or sulfur;<br><br> Y is oxygen or NR6;<br><br> R1 is R7 or R9;<br><br> R2, R3 are independently hydrogen, methyl, -CH2(C6H4)C1. 4alkylene-COOH, -Ci-4alkylene-COO-CH2(C6H5), -Cx_4alkylene-CO-NH-CH2(C6H5), saccharide, -Ci.4alkylene-CH2-NH(2.n)(Ci.4alkyl)n wherein n is 2 or 1;<br><br> R4 is methyl, -CH2(C6H4)Ci.4alkylene-COOH, -Ci_4alkylene-COO-CH2(C6H5), -Ci.4alkylene-CO-NH-CH2 (C6Hs), saccharide, -Q. 4alkylene-CH2-NH(2.n)(Ci.4alkyl)n wherein n is 2 or 1;<br><br> R5 is methyl or R8;<br><br> R6 is hydrogen or methyl<br><br> R7 is -Ci.i0alkylene-COOH, -Ci„4alkylene-CONH2, -Q. 4alkylene-COO-Ci.4alkyl, -Ci.4alkylene-CON(Ci.4alkylene-COOH)2, -Ci.<br><br> INTELLECTU OFRCF<br><br> 23<br><br> RECl<br><br> XL PROPERTY OF N.Z<br><br> \R 2004<br><br> ElVED<br><br> 4alkylene-OH, -Ci.4alkylene-NH3-halo or -Cx.4alkylene-0S02NH(C 4alkyl);<br><br> R8 is -C7.x7alkyl, -COOH, -C7.17 olefinic group containing to 5 ethylenic bonds, -C=C-COO-C1.4alkyl, or -C^alkylene-COO-Ci alkyl;<br><br> R9 is -Ci.4alkylene-0-Cx.4alkyl, -Ci-i0alkylene-CO-SH, -CL 4alkylene-CO-S(C!.4alkyl), -Cx.4alkylene-CS-NH2, saccharide, alkoxy-linked saccharide, -Cx-4alkylene-CO-NH(2-n)(Cx-4alkyl)n wherein n is or 1, -Cx-4alkylene-S02-0(Cx-4alkyl), -C1.4alkylene-0S02-0(C1.4alkyl), Cx-4alkylene-0P(0-Cx-4alkyl)3, or -Cx.10alkylene-CN; and with the proviso that X and Y cannot both be oxygen when<br><br> R1 is R7,<br><br> R2, R3 are independently hydrogen or Ci_4alkyl;<br><br> R4 is Ci_4alkyl; and<br><br> R5 is R8.<br><br> 2. The compound of claim 1, wherein said compoun is l-aza-a-tocopherol-6-yloxyl-acetic acid, 1-aza-a-tocopherol yloxyl-methyl acetate, 1 -aza-N-methyl-a-tocopherol-6-yloxyl-methyl acetate, or l-aza-N-methyl-a-tocopherol-6-yloxyl-acetic acid.<br><br> 3. A compound 6-(2,4-dinitrophenylazo (2,5,7,8-tetr methyl-2-(4,8,12-trimethyltri decyl)-1,2,3,4-tetrahydroquinoline.<br><br> 4. The compound of any of claims 1 to 3, wherein said compound exhibits an antiproliferative effect comprising<br><br> 89<br><br> INTELLECTUAL PROPERTY<br><br> OFPICP OF N.Z<br><br> 2 3 MAR 2M1!<br><br> RECEIVED<br><br> apoptosis, DNA synthesis arrest, cell cycle arrest, or cellular differentiation.<br><br> 7. A pharmaceutical compound comprising the compound of any of claims 1 to 6 and a pharmaceutically acceptable carrier.<br><br> 8. A compound as claimed in any of claims 1 to 3 substantially as hereinbefore described with reference to the examples.<br><br> 9. A pharmaceutical compound comprising the compound of claim 8 and a pharmaceutically acceptable carrier.<br><br> 10. Use of the compound of any of claims 1 to 3 in the preparation of a medicament to treat a cell proliferative disease.<br><br> 11. Use according to claim 10, wherein said cell proliferative disease is a neoplastic disease, a non-neoplastic disease or a non-neoplastic disorder.<br><br> 12. Use according to claim 11, wherein said neoplastic disease is ovarian cancer, cervical cancer, endometrial cancer,<br><br> bladder cancer, lung cancer, breast cancer, testicular cancer,<br><br> prostate cancer, gliomas, fibrosarcomas, retinoblastomas,<br><br> melanomas, soft tissue sarcomas, ostersarcomas, leukemias, colon<br><br> 90<br><br> INTELLECTUAL PROPERTY<br><br> OFFICE OF N.Z<br><br> 2 3 MAR 20M<br><br> RECEIVED<br><br> cancer, carcinoma of the kidney, pancreatic cancer, basal cell carcinoma, or squamous cell carcinoma.<br><br> 13. Use according to claim 11, wherein said non neoplastic disease is selected from the group consisting of psoriasis(, benign proliferative skin diseases, ichthyosis, papilloma, restinosis scleroderma, hemangioma, viral diseases, and autoimmune diseases<br><br> 14. Use according to claim 13, wherein said autoimmune diseases are selected from the group consisting o autoimmune thyroiditis, multiple sclerosis, myasthenia gravis;, systemic lupus erythematosus, dermatitis herpetiformis, celiac disease, and rheumatoid arthritis.<br><br> 15. Use according to claim 11, wherein said non neoplastic disorder is a viral disorder or an autoimmune disorder.<br><br> 16. Use according to claim 15, wherein said vircjl disorder is Human Immunodeficiency Virus.<br><br> 17. Use according to claim 15, wherein said autoimmune disorder is the inflammatory process involved in cardiovascular plaque formation, ultraviolet radiation induced skin damage or a disorder involving an immune component.<br><br> end of claims<br><br> 91<br><br> INTELLECTUAL PROPERTY<br><br> OFHCF OF N.Z<br><br> 23 MAR 200*1<br><br> </p> </div>
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Families Citing this family (38)

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Publication number Priority date Publication date Assignee Title
US6703384B2 (en) 1998-09-23 2004-03-09 Research Development Foundation Tocopherols, tocotrienols, other chroman and side chain derivatives and uses thereof
CA2399802C (en) 2000-02-11 2009-12-08 Research Development Foundation Tocopherols, tocotrienols, other chroman and side chain derivatives and uses thereof
JP2004513183A (en) * 2000-11-14 2004-04-30 バイタル　ヘルス　サイエンシズ　プロプライアタリー　リミティド Complex of phosphoric acid derivative
AUPR549901A0 (en) * 2001-06-06 2001-07-12 Vital Health Sciences Pty Ltd Topical formulation containing tocopheryl phosphates
JP4745608B2 (en) * 2001-07-27 2011-08-10 バイタル ヘルス サイエンシズ プロプライアタリー リミティド Skin treatment using phosphoric acid derivatives of electron transfer agents
AU2002350272B1 (en) * 2001-12-13 2003-06-23 Vital Health Sciences Pty Ltd Transdermal transport of compounds
US20030236301A1 (en) * 2001-12-19 2003-12-25 Bob Sanders Liposomal delivery of vitamin E based compounds
AU2002950713A0 (en) 2002-08-09 2002-09-12 Vital Health Sciences Pty Ltd Carrier
JP4690305B2 (en) * 2003-01-17 2011-06-01 バイタル ヘルス サイエンシズ プロプライアタリー リミティド Compounds with antiproliferative properties
AU2003901815A0 (en) * 2003-04-15 2003-05-01 Vital Health Sciences Pty Ltd Phosphate derivatives
GB0313612D0 (en) 2003-06-12 2003-07-16 Novartis Ag Organic compounds
EP1664067A2 (en) * 2003-09-15 2006-06-07 DSM IP Assets B.V. A NEW ROUTE TO alpha-TOCOPHEROL, alpha-TOCOPHERYL ALKANOATES AND PRECURSORS THEREOF
WO2005084678A1 (en) * 2004-03-03 2005-09-15 Vital Health Sciences Pty Ltd Alkaloid formulations
EP1858508A4 (en) * 2005-03-03 2009-01-07 Vital Health Sciences Pty Ltd Compounds having anti-cancer properties
CA2611831C (en) 2005-06-17 2014-09-16 Vital Health Sciences Pty Ltd. A carrier comprising one or more di and/or mono-(electron transfer agent) phosphate derivatives or complexes thereof
EP1973547A1 (en) * 2005-12-23 2008-10-01 Vital Health Sciences Pty Ltd. Compounds having cytokine modulating properties
JP5256049B2 (en) * 2006-02-21 2013-08-07 ザ オハイオ ステート ユニバーシティー リサーチ ファウンデーション Anticancer drug
US8318812B2 (en) 2006-06-02 2012-11-27 The Ohio State University Research Foundation Therapeutic agents for the treatment of lymphoid malignancies
US8288369B2 (en) * 2006-06-27 2012-10-16 University Of South Florida Delta-tocotrienol treatment and prevention of pancreatic cancer
WO2009012096A2 (en) * 2007-07-18 2009-01-22 Research Development Foundation Improved therapeutic methods and compositions comprising chroman ring compounds
US8598145B2 (en) 2008-03-14 2013-12-03 Stephen John Ralph Mitochondrially delivered anti-cancer compounds
US8461362B2 (en) 2009-04-13 2013-06-11 The Ohio State University Research Foundation Protein phosphatase 2A-activating agents
CA2759011A1 (en) 2009-04-17 2010-10-21 The Ohio State University Research Foundation Antiadhesion agents
US10071030B2 (en) 2010-02-05 2018-09-11 Phosphagenics Limited Carrier comprising non-neutralised tocopheryl phosphate
ES2829386T3 (en) 2010-03-30 2021-05-31 Phosphagenics Ltd Transdermal administration patch
US8309768B2 (en) 2010-11-29 2012-11-13 The Ohio State University Research Foundation FTY720-derived anticancer agents
EP2685992A4 (en) 2011-03-15 2014-09-10 Phosphagenics Ltd Amino-quinolines as kinase inhibitors
RU2613110C2 (en) * 2012-05-18 2017-03-15 Те Огайо Стейт Юниверсити Methods for improving liver function
JP2015518896A (en) * 2012-06-06 2015-07-06 ユニザー ヴィロロジー，エルエルシー Novel imino sugars and their uses
UA119740C2 (en) * 2012-06-13 2019-08-12 Сайтек Текнолоджи Корп. Stabilizer compositions containing substituted chroman compounds and methods of use
WO2015035212A1 (en) * 2013-09-06 2015-03-12 Research Development Foundation Alpha-tea salt forms: compositions and uses for treating disease
BR112016005509B1 (en) 2013-09-13 2021-01-19 The Board Of Trustees Of The University Of Arkansas use of a composition
US20160324914A1 (en) 2015-05-05 2016-11-10 Tocol Pharmaceuticals, Llc Use of rice bran oil distillate extract for prevention and mitigation of the effects of radiation
CN105078963B (en) * 2015-09-29 2018-06-22 中山大学 Alpha-tocopherol is preparing the application in treating snail fever drug
AU2016367708B2 (en) 2015-12-09 2022-07-14 Phosphagenics Limited Pharmaceutical formulation
MX2019006845A (en) 2016-12-21 2019-10-15 Avecho Biotechnology Ltd Process.
KR20180002728U (en) 2017-03-10 2018-09-19 주식회사 두울온돌침대 Mattress with electric heat
CA3135920A1 (en) * 2020-10-28 2022-04-28 Hermay Labs Corporation Labile and coherent redox-silent analogues for vitamin e enhancement

Family Cites Families (57)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5118954B2 (en) * 1972-02-04 1976-06-14
CS172015B1 (en) 1974-05-12 1976-12-29
US4181725A (en) * 1977-05-02 1980-01-01 The Regents Of The University Of Michigan Method for alleviating psoriasis
US4320141A (en) * 1977-12-16 1982-03-16 Yasuhiro Komatsu Antitumor agent
JPS6056155B2 (en) 1978-12-04 1985-12-09 エーザイ株式会社 Chromanyloxy fatty acid congeners
US4457918A (en) * 1982-05-12 1984-07-03 The General Hospital Corporation Glycosides of vitamins A, E and K
DE3309159A1 (en) * 1983-03-15 1984-09-20 Basf Ag, 6700 Ludwigshafen NEW OPTICALLY ACTIVE CHROME DERIVATIVES, METHOD FOR THE PRODUCTION THEREOF AND NEW INTERMEDIATE PRODUCTS
US4873088A (en) * 1983-09-06 1989-10-10 Liposome Technology, Inc. Liposome drug delivery method and composition
US4617317A (en) * 1984-10-31 1986-10-14 Bennet Justin D Method of treating ulcerative colitis
DE3686025T2 (en) * 1985-05-22 1993-01-07 Liposome Technology Inc METHOD AND SYSTEM FOR INHALING LIPOSOMES.
US4665204A (en) * 1985-06-27 1987-05-12 Henkel Corporation Diester derivatives of tocopherol
JPS63303049A (en) 1987-06-04 1988-12-09 Shinagawa Refract Co Ltd Fiber-reinforced vitreous thermal spraying material
JPS6441727A (en) 1987-08-07 1989-02-14 Toshiba Corp Cooking unit
US5610180A (en) * 1988-01-29 1997-03-11 Virginia Commonwealth University Ionizable congeners of aromatic and aliphatic alcohols as anti-leukemia agents
US5198432A (en) * 1988-01-29 1993-03-30 Center For Innovative Technology Method of preventing chlorohydrocarbon toxicity using sterol derivatives
US5260294A (en) * 1988-08-09 1993-11-09 Hoffman-La Roche Inc. Chromanes and their pharmaceutical compositions and methods
JPH02215778A (en) * 1989-02-14 1990-08-28 Kuraray Co Ltd 3,4-dihydro-2h-benzopyrane derivative and medical use of the same compound
JPH03206089A (en) 1990-01-05 1991-09-09 Eisai Co Ltd New vitamin e derivative and production thereof
DE4014708C2 (en) 1990-05-08 1994-03-10 Heidelberger Druckmasch Ag Procedure for determining register errors
US5114957A (en) * 1990-05-08 1992-05-19 Biodor U.S. Holding Tocopherol-based antiviral agents and method of using same
WO1993009777A1 (en) * 1991-11-22 1993-05-27 Lipogenics, Inc. Tocotrienols and tocotrienol-like compounds and methods for their use
FR2672601B1 (en) 1991-02-08 1994-10-14 Synthelabo BENZO-1,5-THIAZEPINE DERIVATIVES, THEIR PREPARATION AND THEIR THERAPEUTIC APPLICATION.
JPH05213982A (en) 1991-03-26 1993-08-24 Dainippon Ink & Chem Inc Oligosaccharide tocopherol glycoside, sulfated oligosaccharide tocopherol glycoside and antiviral agent comprising the same as active ingredient
FR2676056B1 (en) * 1991-05-03 1993-07-16 Adir NOVEL BENZOPYRAN DERIVATIVES, THEIR PREPARATION PROCESS AND THE PHARMACEUTICAL COMPOSITIONS CONTAINING THEM.
EP0636618A1 (en) 1991-06-04 1995-02-01 Marigen S.A. Spontaneously dispersible concentrates and microemulsions with vitamin E compounds for the preparation of medicaments with anti-tumour activity
US5135945A (en) * 1991-06-05 1992-08-04 Merrell Dow Pharmaceuticals Inc. Cholesterol-lowering tocopherol analogs
US5545660A (en) * 1992-04-07 1996-08-13 Merrell Pharmaceuticals Inc. Hydrazide derivatives of 3,4-dihydro-2H-1-benzopyrans
DE4215536A1 (en) * 1992-05-12 1993-11-18 Basf Ag Cyan mixtures for dye transfer
FR2695387B1 (en) * 1992-09-09 1994-10-21 Adir New benzopyran compounds, process for their preparation and pharmaceutical compositions containing them.
US5403834A (en) * 1992-12-07 1995-04-04 Eukarion, Inc. Synthetic catalytic free radical scavengers useful as antioxidants for prevention and therapy of disease
IT1256264B (en) * 1992-12-31 1995-11-29 Lifegroup Spa N-HYDROXYAMINE N-ACIL DERIVATIVES EQUIPPED WITH SCAVENGER ACTIVITIES AND USABLE IN ACUTE AND CHRONIC PATHOLOGIES RELATED TO PHENOMENA OF PEROXIDATION AND INFLAMMATION
DE4304303C2 (en) 1993-01-22 1997-01-30 Pacific Corp Vitamin E derivatives with a quaternary nitrogen atom, process for their preparation and cosmetic compositions containing the same
CA2152693A1 (en) * 1994-08-05 1996-02-06 Kazumi Ogata Therapeutic composition for pancreatitis
US5670255A (en) * 1995-01-23 1997-09-23 Ppg Industries, Inc. Antioxidant compositions for coating substrates, substrates coated with the same and methods for inhibiting the oxidation of such compositions applied to a substrate
DE19504398A1 (en) * 1995-02-10 1996-08-14 Beiersdorf Ag Tocopherylglycoside, their preparation and their use as surfactants, as antioxidants and as a cell aging preventive agent in cosmetic or pharmaceutical preparations
DE19523079A1 (en) 1995-06-26 1997-01-02 Basf Ag Esters and amides of 9 (Z) -retinoic acid
CA2179940A1 (en) 1995-07-18 1997-01-19 Kazumi Ogata Antiallergic composition comprising a phosphoric diester compound
US5798356A (en) 1995-08-07 1998-08-25 Alcon Laboratories, Inc. Angiostatic compounds
CA2186654A1 (en) 1995-10-13 1997-04-14 Masayuki Nakamura Phosphoric diester
JPH09301953A (en) * 1996-03-12 1997-11-25 Sanwa Kagaku Kenkyusho Co Ltd Malonic acid diamide derivative and use thereof
JPH104489A (en) * 1996-04-19 1998-01-06 Nikon Corp Picture input device
JPH09301969A (en) * 1996-05-15 1997-11-25 Fuji Photo Film Co Ltd Chroman-based compound and heat-sensitive recording material using the same
DK0815858T3 (en) * 1996-07-01 2001-10-08 Nisshin Flour Milling Co Pharmaceutical composition containing 9-cis-retinoic acid alpha-tocopherol ester for the treatment of leukemia
US6022560A (en) * 1996-08-27 2000-02-08 Clarion Pharmaceuticals Inc. Pharmaceutical compositions, novel uses, and novel form of α-tocopherylphosphocholine
FR2754713B1 (en) 1996-10-22 1999-01-08 Roc Sa USE OF COMPLEXES FOR THE PREPARATION OF COMPOSITIONS FOR THE TREATMENT OF SENSITIVE SKIN, PREPARATION METHOD AND HYPOALLERGENIC COMPOSITIONS
JP2001507363A (en) * 1997-03-04 2001-06-05 ウイスコンシン アラムナイ リサーチ フオンデーシヨン Method for suppressing tumor growth by combining isoprenoids and statins
CA2220541A1 (en) 1997-11-07 1999-05-07 Mcgill University Analogs of vitamin e
US5917060A (en) * 1997-12-17 1999-06-29 Basf Aktiengesellschaft Preparation of chromanyl derivatives
US6770672B1 (en) * 1998-09-23 2004-08-03 Research Development Foundation Tocopherols, tocotrienols, other chroman and side chain derivatives and uses thereof
US6703384B2 (en) * 1998-09-23 2004-03-09 Research Development Foundation Tocopherols, tocotrienols, other chroman and side chain derivatives and uses thereof
RU2232758C2 (en) * 1998-09-23 2004-07-20 Рисерч Дивелопмент Фаундейшн Tocopherols, tocotrienols and other derivatives of chroman and by-side chains and methods for treatment with their using
FR2784988B1 (en) * 1998-10-23 2002-09-20 Adir NOVEL DIHYDRO AND TETRAHYDROQUINOLEINIC COMPOUNDS, PROCESS FOR THEIR PREPARATION AND THE PHARMACEUTICAL COMPOSITIONS CONTAINING THEM
US5990322A (en) * 1999-03-09 1999-11-23 Sk Corporation Alpha-tocopherol cyclopropylates, the new vitamin E derivatives and method for producing the same
US6127382A (en) * 1999-08-16 2000-10-03 Allergan Sales, Inc. Amines substituted with a tetrahydroquinolinyl group an aryl or heteroaryl group and an alkyl group, having retinoid-like biological activity
CA2399802C (en) * 2000-02-11 2009-12-08 Research Development Foundation Tocopherols, tocotrienols, other chroman and side chain derivatives and uses thereof
US20030236301A1 (en) * 2001-12-19 2003-12-25 Bob Sanders Liposomal delivery of vitamin E based compounds
WO2003089613A2 (en) * 2002-04-19 2003-10-30 Research Development Foundation Tocopherol associated protein and uses thereof

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